[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

AU681998B2 - Marcfortine/paraherquamide derivatives useful as antiparasitic agents - Google Patents

Marcfortine/paraherquamide derivatives useful as antiparasitic agents Download PDF

Info

Publication number
AU681998B2
AU681998B2 AU31264/93A AU3126493A AU681998B2 AU 681998 B2 AU681998 B2 AU 681998B2 AU 31264/93 A AU31264/93 A AU 31264/93A AU 3126493 A AU3126493 A AU 3126493A AU 681998 B2 AU681998 B2 AU 681998B2
Authority
AU
Australia
Prior art keywords
marcfortine
alkyl
thiomarcfortine
alkoxy
benzyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
AU31264/93A
Other versions
AU3126493A (en
Inventor
Byung Hyun Lee
Vincent P. Marshall
Stephen J Nelson
Renate N Taylor
Howard A Whaley
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pharmacia and Upjohn Co
Original Assignee
Pharmacia and Upjohn Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Pharmacia and Upjohn Co filed Critical Pharmacia and Upjohn Co
Publication of AU3126493A publication Critical patent/AU3126493A/en
Assigned to PHARMACIA & UPJOHN COMPANY reassignment PHARMACIA & UPJOHN COMPANY Amend patent request/document other than specification (104) Assignors: UPJOHN COMPANY, THE
Application granted granted Critical
Publication of AU681998B2 publication Critical patent/AU681998B2/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D491/00Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/22Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains four or more hetero rings
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/90Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00Antiparasitic agents
    • A61P33/10Anthelmintics
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/18Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Virology (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Agronomy & Crop Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Environmental Sciences (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Pyridine Compounds (AREA)

Description

COR~RETED
'I RSION
*I
Pcr 3 q -t3 page 46, description, replaced by new page 46 WORLD INTELLECTUAL PROPERTY ORGANIZATION International Bureau 0 INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (51) International Patent Classification 5 (11) International Publication Number: WO 93/10120 C07D 491/22, A01N 43/90 C2P 17/18, C12N 1/14 A61K 31/495 (C07D 491/22 Al C07D 321:00, 241:00, 221:00 C07D 221:00, 209:00) (43) International Publication Date: 27 May 1993 (27.05,93) (C12P 17/18, C12R 1:00) (21) International Application Number: PCT/US92/09483 (72) Inventors; and Inventors/Applicants (for US only) LEE, Byung, Hyun (22) International Filing Date: 13 November 1992 (13.11.92) [KR/US]; 5905 Stoney Brook, Kalamazoo, MI 49002 TAYLOR, Renate, N. [US/US]; 2401 Fairgrove Priority data: Street, Kalamazoo, MI 49009 WHALEY, Howard, 795,457 22 November 1991 (22.11.91) US A. [US/US]; 5619 Stonebrook, Portage, MI 49002 (US).
795,900 22 November 1991 (22.11.91) US NELSON, Stephen, J. [US/US]; 6540 South 42nd 796,948 22 November 1991 (22.11.91) US Street, Climax, MI 49034 MARSHALL, Vincent, 797,736 22 November 1991 (22.11.91) US P. [US/US]; 204 Paisley Court, Kalamazoo, MI 49006
(US).
Parent Applications or Grants (63) Related by Continuation (74) Agent: JAMESON, William, Corporate Intellectual US 07/795,457 (CIP) Property Law, The Upjohn Company, 301 Henrietta Filed on 22 November 1991 (22.11.91) Street, Kalamazoo, MI 49001 (US).
US 07/795,900 (CIP) Filed on 22 November 1991 (22.11.91) (81) Designated States: AU, BB, BG, BR, CA, CS, FI, HU, JP, US 07/796,948 (CIP) KP, KR, LK, MG, MN, MW, NO, PL, RO, RU, SD, Filed on 22 November 1991 (22.11.91) UA, US, European patent (AT, BE, CH, DE, DK, ES, US 07/797,736 (CIP) FR, GB, GR, IE, IT, LU, MC, NL, SE), OAPI patent Filed on 22 November 1991 (22.11.91) (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, SN, TD, t'+ Q.t' 6\ i TG).
(71) Applicant (for a estgaed St exept U TG).
JOHN-GOM4PARY- [US/US]; 301 Henrietta Street, Kal- Published amazoo, MI 49001 With international search report.
Before the expiration of the time litit for amending the 6 A8 A claims and to be republished in the event of the receipt of (54) Title: MARCFORINE/PARAHERQUMIDE DERIVATIVES USEFUL AS ANTIPARASITIC AGENTSndmen (54)Title: MARCFORTINE/PARAHERQUM IDE DERIVATIVES USEFUL AS ANTIPARASITIC AGENTS (57) Abstract There are disclosed 18-thiomarcfortine derivatives of the natural products marcfortine A, B, and C, C-18 thioparaherquamide and derivatives thereof, novel N-l marcfortines A, B, and C and derivatives thereof, novel N- paraherquamide and derivatives thereof useful in the treatment and prevention of helminth and arthropod infections of animals and plants. The synthetic derivatives are of Formula (Referred to in PCT Gazette No. 2111993, Section II) 'WO 93/10120 PCT/US92/09483 MARCFORTINE/PARAHERQUAMIDE DERIVATIVES USEFUL AS ANTIPARASITIC AGENTS BACKGROUND OF THE INVENTION The marcfortines are known compounds and are disclosed by Polonsky et al. in Journal of the Chemical Society Chemical Communications 1980 601-602 (Marcfortine A) and Tetrahedron Letters 1981 22 1977-1980 (Marcfortines B and The compounds are fungal metabolites of Penicillium roqueforti. The marcfortines are structurally related to the paraherquamides which are also known compounds. The paraherquamides are disclosed by Yamazaki et al. in Tetrahedron Letters 1981 22 135-136, and by Blanchflower et al., Journal of Antibiotics, 1991, 44, 492-497. U.S. Patents 4,866,060 and 4,923,867 disclose the use of the marcfortines A, B, and C, and certain derivatives thereof as useful for the treatment and prevention of parasitic diseases in animals.
Paraherquamide has the following structure:
H
3 C ,CH 3 0 7 24 16 H 3 C CH 3 12> 3 14 13 21 3 INH
H
3 C ,19 0H 20
N
Marcfortine A has the following structure: WO 93/10120 WO 9310120PCT/US92/09483'.
-2- Marcfortine B has the following structure: 61 2 524 Marcfortine, C has the following structure:
,CH
3 2 WO 91/09961 (published 11 July 199 1) discloses various derivatives of marcfortine and paraherquamnide, and N(12) oxides thereof, well as the production the production of VM 29919 (paraherquamide) and VM 55596 (the N(12)oxide of paraherquamide) inter alia from Penicillium Sp. INiL 332995.
US Patent 4,873,247 discloses derivatives of paraherquamnide and a strain of Penicilum ,,harlessi MW 5123 (ATCC 20841) for the production of paraherquamnide. US Patent 4,978, 656 (as well as EP 390532-A, EP-301 742-A) discloses various synthetic derivatives of paraherquarnide as well as the production of paraherquamide from Penicillium charlessi NT 5123 (ATCC 20841).
Paraherquaniide is a compound produced under certain conditions by the fungal organism Penicilium paraherquei. WO 92/00300 (published 9 January 1992) discloses the WO 93/10120 PCT/US92/09483 -3synthesis paraherquamide related compounds produced from Penicillium paraherquei, deposited at the C.A.B. International Mycological Institute, Ferry Lane, Kew, London, under deposit number CMI 68220.
SUMMARY OF THE INVENTION This invention is concerned with the synthesis of C-18 thiomarcfortines A. B, and C and derivatives thereof, C-18 thioparaherquamide and derivatives thereof, novel N-1 marcfortines A, B, and C and derivatives thereof, novel N-1 paraherquamide and derivatives thereof, and the use of these compounds as antiparasitic agents. Thus it is an object of this invention to describe these thiomarcfortine, thioparaherquamide, marcfortine and paraherquamide derivatives. A further objective of this invention is to describe processes for the preparation of these compounds. A still further objective is to describe the use of the instant compounds as antiparasitic agents in the treatment and prevention of parasitic diseases. A still further object is to describe compositions for the treatment of parasitic diseases which contain the novel compounds of this invention as the active ingredient thereof. Further objectives will become apparent from a reading of the following description.
DESCRIPTION OF THE INVENTION The compounds of the instant invention are represented by Formula I: H3C CFH3 2 (0)m 0R 2 C n II3 C H 3 7 0 R24 16 N 21'- 1 1413 2 -R, 610 0 R4a Rl4b 18 W R18 a wherein: n is 0 or 1; Ri 4 a and R14b, being the same or different, are selected from hydrogen, hydroxy, Ci-C 6 alkyl, Ci-C 6 alkoxy, Ci-C 6 alkenyl, Ci-C 6 alkenyl-Ci-C 6 alkoxy, CI-C 6 alkynyl-Ci-C 6 alkoxy,
CI-C
6 alkynoyloxy, poly C-C 6 alkoxy-C-C, alkoxy, phenyl, phenyl-C-C, alkyl, tri-Ci-C 6 alkylsilyloxy, diphenylphosphoryloxy and halogen, or Ri 4 a and R1 4 b together form the epoxide or
=CH
2 with the provisio that when n is 1, then R, 4 and RI 4 b are both hydrogen; m is 0 or 1 (preferably 0); WO 93/10120 WO 9310120PCT/US92/09483 -4- W is 0 or S; when W is S, R, is hydrogen, CI-C 7 alkyl, cyclo C 3 -Cgalkyl, benzyl, C 2 -C7 alkanoyl (-C(0)C 2
-C
7 alkyI) (optionally substituted with carboxy (-COOH), C,-0 7 alkanoyl, carbo
C
1
-C
7 alkoxy (-C(O)0C,-C 7 alkyl), -NR 4
R
5 aminocarbonyl (-C(0)NR 4
R
5
C
10
-C
2 4 alkanoyl Io)C 1 4 aikY], cyclo C 3 -Csalkanoyl [optionally substituted with carboxy, C 1
C
7 alkanoyl, carboC,-Cqalkoxy, -NR 4
R
5 aminocarbonyl), alkanoyloxymethylene (-CH 2 OC(0)-C 2
-C
7 alkyl), benzoyloxymethlene (-CH 2 OC(O)phenyl) [optionally substituted with 1 or 2 groups selected from halogen, C,-C 4 alkyl, ha~oC,-C 7 alkyl, nitro, cyano and C 1
-C
7 adkoxy}, C 10
-C
24 alkenoyl
(-C(O)C
9 -C~alkenyl), benzenesulfonyl (-SOCH~phenyl) (optionaliy substiiuted with 1 or 2 groups selecte.d from halogen, C,-C 4 alkyl, ha~oC 1
-C.
7 alkyl, nitro, cyano and Cl-C 7 alkoxy),
C
1 -C~alkylaminocarbonyl (-C(0)N(Cj-C 4 alkyl) 2
C
1
-C
4 all~ylamiriothiocarbonyl (-C(S)N
(CI-C
4 alkyl) 2
C
1
-C
7 alkoxycarbonyl, phenoxycarbonyl (optionally substituted with I or 2 groups selected from halogen, CI-C 4 alkyl, ha~oC,-C~alkyl, nitro, cyano and Cl-C 7 alkoxy}, -C(0)NR' 4 R's, 2
X(R
5
-SR
6
-SO
2
NR
4 R5, benzoyl substituted at the 3 or 4 position with
-CH
2
NR
4
R
5 or 2-tetrahydrofuran or bicycloC 8
-C
12 alkanoyJ; when W is 0; R, is selected from the group consisting of:
C
2
-C
7 alkanoyl substituted with carboxy (-COOH), C,-C 7 alkanoyl, carboC 1
-C
7 alkoxy (-C(0)0C,-C 7 alkyl), -NR 4 aminocarbonyl
(-C(O)NR
4
R
5 cyclo C 3 -Calkanoyl optionally substituted with carboxy, C,-C 7 alkanoyl, carboC,-C 7 alkoxy, -NR 4
R
5 aminocarbonyl; alkanoyloxymethylene (-CH 2 OC(0)-C 2
-C
7 alkyl); benzoyloxymethlene (-CH 2 OC(0)phenyl) substituted with 1 or 2 groups selected from halogen, C,-C 4 alkyl, haloC,-C~alkyl, nitro, cyano and C,-C~alkoxy; phenoxycarbonyl substituted with I or 2 groups selected from halogen,
C,-C
4 alkyl, haloC,-C~alkyl, nitro, cyano and CI-C 7 alkoxy;
-C(O)NR'
4
R'
5 2
)(R
3 -SR6.
Cl 0 -C4alkenoyl (-C(0)C 9
-C
23 alkenyl); or 2-tetrahydrofuran;
R
4 and R 5 being the same or different, are selected from hydrogen, C 1
-C
7 alkyl, phenyl (optionally substituted with 1 or 2 groups selected from halogen, C 1
-C
4 alkyl, haloC,-C~alkyl, nitro, cyano and C,-C 7 alkoxy} or when taken together with N, form a saturated WO 93/10120 WO 9310120PCT/US92/09483 or unsaturated heterocyclic amine ring;
R'
4 and R' 5 being the same or different, are selected from C,-C7 alkyl, cyclo(C 3
-C
8 )alkyl, phenyl {optionally substituted with I or 2 groups selected from halogen,
C,-C
4 alkyl, haloCI-C 7 alkyI, nitro, cyano and C,-C 7 alkoxy] or when taken together with N, form a saturated heterocyclic amnine ring optionally containing I or 2 additional heteroatoms selected from N, 0 or S; X is 0 or S;
R
2 and R 3 being he same or different, are selected from C,-C 7 alkyl, phenyl {optionally substituted with 1 or 2 groups seiected from halogen, alkyl, haloC,-C 7 alkyl, nitro, cyano and C 1
-C
7 alkoxyj, C 1 alkoxy, thio(C 1
-C
1 )alkoxy, phenoxy, Nhophenoxy,
-NR
7 R, f{where R 7 and being the same or different, are selected from H, CX- 7 alkyl or taken toglether with N, form a satural-od heterocyclic ring), or taken together with P form a 4- to 7-membered heterocyclic ring; RO s C,-C7 alkyl, haloCI-C~alkyl, carboCI-C 7 alkoxy, -NR 9
R
1 0 where R 9 and RIO, being the sanm or different, are C,-C 7 alkyl or phenyl (optionally substituted with I or 2 groups selected from halo, lower alkyl, haloC 1
C
7 alkyl, nitro, cyalio, C 1
-C
7 alkoxy); R24 is hydrogen, halogen or C,-C 7 alkoxy; is hydrogen or halogen; is hydrogen, C,-C 7 alkyl, alkoxyalkyl, C 2
-C
8 alkenyl, C 2 -Cs alkynyl or benzyl; the broken line between carbons 24 and 25 represents a single or double bond.
The compounds of this invention include pharmaceutically acceptable salts thereof as well as !2a-N-oxides thereof.
Another aspect of this invention are 10,1 1,4-dicarbonylpiperazine) 10', 10'a-hexahydro- 12'pentainethyl)- [2'S-[2'.alpha.,3'a~apha., 9'a.alpha., 3"'aS*9"aS*, 1'abeta.]J-Spiro[4H,8H-[1I,4]dioxepino[2,3-g]indole-8,2'(3'H)- I H,4-- 3a,9a1(iminomethano) cyclopenta[biquinolizinl-9,i l'(IOH)-dione Cpd #120), and 10, 10"-(1I,4-dioxo-2-butene)bis(6',7',8',9', 10', l0'a-hexahydro- 12'-pentamethyl)- [2'S-j12'.alpha.,3'a.alpha.,9'a.alpha.,1I 10'a.beta.1- Spiro[4H,8H-[1 ,4ldioxepino[2,3-glindole-8,2'(3'H)-[ IH,4H-3a,9a](imino methano)cyclopentabl quinolizinl-9,1 1(10H)-dione, Cpd #1 2T), which can be used in the same manner as compounds of Formula L.
WO 93/10120 WO 9310120PCT/US92/09483 -6- Another aspw~t of this invention provides an 18-thiomarcfortine or derivatives of Formula 11: fl 3 C %Cl 6 5,R 2 fl 3 C H ,-2 16 N 1 113
R
"19 10 0 18
N\
R8 wherein: m is 0 or I (preferably 0); R, is hydrogen, CI-C 7 alkyl, cyclo C 3 -Csalkyl, benzyl, q2-C7 alkanoyl
(-C(O)C
2 -C~alkyl) (optionally substituted with carboxy (-COOH), C,-C 7 alkanoyl, carbo
C
1
-C
7 alkoxy (-C(O)OCI-Cqalkyl), -NR 4
R
5 aminocarbonyl (-C(O)NR 4
R
5
C
10
-C
2 4 alkanoyl
(-C(O)CI
0
-C
24 alkyl, cyclo C 3 -Csalkanoyl (optionally substituted widi carboxy, Cl-C? alkanoyl, carboC,-C~alkoxy, -NR 4 Rs, aminocarbonyl I, alkanoyloxymethylene (-CH 2
QC(O)-C
2 -C~alkyl), benzoyloxymethlene (-CH 2 OC(O)phenyl) f(optionally substituted with 1 or 2 groups selected from hpa!ogen, C,-C 4 alkyl, haloC,-Cqalkyl, nitro, cyano and C 1 -C~alkoxyj, CI 0 -Cq 4 alkenoyl
(-C(O)C
9
-C,
3 alkenyl), benzenesulfr-nyl (-SO 2
CH
2 phenyl) (optionally substituted with 1 or 2 groups selected from halogen, C,-C 4 alkyl, haloC,-C~alkyl, nitro, cyano and C 1
-C
7 alkoxy), CI-C 4 alkylaminocarbonyl (-C(O)N(C 1
-C
4 alkYl) 2 Cl-C 4 alkylamninothiocarbonyl 4 alkyI) 2
CI-C
7 alkoxycarbony!, phenoxycarbonyl (optionally substituted with 1 or 2 groups selected from halogen, CI-C 4 alkyl, haloC,-C~alkyl, nitro, cyano and C,-C~alkoxy), -C(O)NR' 4
R'
5 2
(R
3
-SR
6
-SO
2
NR
4 R5, benzoyl substituted at the 3 or 4 position with -CH 2
NR
4
R,
2-tetrahydrofuran, or bicycloC.-C 12 alkanoyl;
R
4 and R 5 being the same or different, are selected from hydrogen, CI-C7 alkyl, phenyl (optionally substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, halo C,-C 7 alkyl, nitro, cyano and CI-C~alkoxy) or when taken together with N, form a saturated or unsaturated heterocyclic amine ring;
R'
4 and R' 5 being the same or different, are selected from C 1
-C
7 alkyl, cyclo(C 3 -Cg)alkyl, phenyl (optionally substituted with 1 or 2 groups selected from halogen,
C,-C
4 alkyl, haloCI-C~alkyl, nitro, cyano and C 1 -C~alkoxy) or when taken together with N, form a saturated heterocyclic amine ring optionally containing I or 2 additional beteroatoms selected from N,0or S; X isO0 or S; WO 93/10120 WO 9310120PCT/US92/09483 -7- R, and R 3 being the same or different, are selected from C,-C 7 alkyl, phenyl loptionally substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, ha~oC 1
-C
7 alkyl, nitro, cyano and Cj-C 7 alkoxy), C 1 -C7 alkoxy, thio(C,-C 7 )alkoxy, phenoxy, thiophenoxy,
-NR
7 R, (where R 7 and being the same or different, are selected from H, Cj-C 7 alkyl or taken together with N, form a saturated heterocyclic ring)I, or taken together with P form a 4- to 7-membered heterocyclic ring;
R
6 is C,-C7 alkyl, haloC,-C 7 alkyl, carboC 1
-C
7 alkoxy, -NR 9 Rj 0 where R 9 and RIO, being the same or different, are Cj-C 7 alkyl or phenyl (optionally substituted with I or 2 groups selected from halo, lower alkyl, haloC,-C 7 alkyl, nitro, cyano, C 1 alkoxy);
R
24 is hydrogen, halogen or C,-C7 alkoxy; is hydrogen or halogen; is hydrogen, Cj-C 7 alkyl, C 2
-C
8 alkoxyalkyl, C 2
-C
8 alkenyl, C 2
-C
8 alkynyl or benzyl; the broken line between carbons 24 and 25 represents a single or double bond.
Another aspect of this invention provides an 18-thioparaherquamide or derivatives of Formula III: 1130 ,013 (O)m 0\ 251 11 2 16 I \113 6 k 14 13 1 11R 19 1 10 0 R14 R4b 8 N1 S R18a wherein: m is 0 or 1 (preferably 0); R, is hydrogen, C,-C 7 alkyl, cyclo C 3 C~alkyl, benzy](2-C alkanoyl 7 a~kyl) (optionally substituted with carboxy (-COOH), C 1 -C7 alkanoyl, carbo Cj-G 7 alkoxy (-C(O)QC 1 -C~alkyl), -NR 4
R
5 aminocarbonyl (-C(O)NR 4 R5)1, CO'C2C1alkanoyl
(-C(O)CI
0 -C24alkyI, cyclo C 3 -C~alkanoyl (optionally substituted with carboxy, CC 7 alkanoyl, carboC,-C~alkoxy, -NR 4
R
5 aminocarbonyl), alkanoyloxym ethylene (-CH 2
OC(O)-C
2
-C
7 alkyl), benzoyloxymethlene (-CH 2 OC(O)phenyl) (optionally substituted with 1 or 2 groups selected from halogen, C 1
-C
4 alkyl, haloC 1 -C~alkyl, nitro, cyano and C,-C 7 alkoxyl, C 10
-C
2 4 alkenoyl WO 93/10120 WO 9310120PCVU1S92/09483 -8-
(-C(O)C
9
-C
23 alkeny)), benzenesulfonyl (-SO 2
CH
2 phenyl) I optionally substituted with 1 or 2 groups selected from halogen, alkyl, haloC,-C 7 alkyl, nitro, cyano and C 1
-C
7 alkoxy),
C
1
-C
4 alkylam inocarbonyl (-C(O)N(C 1
-C
4 aikyl) 2
C
1
-C
4 alkyl aminothiocarbonyl -(N (Cj-C,,alkyl) 2 Cj-C7 alkoxycarbonyl, phenoxycarbonyl [optionally substituted with 1 2 groups selected from halogen, C,-C 4 alkyl, haloC 1
C
7 alkyl, nitro, cyano and C 1
-C
7 alkoxy},
-C,(O)NR'
4
R'
5 2
)(R
3
-SR
6
-SO
2
NR
4 R5, benzoyl substituted at the 3 or 4 position with
-C.H
2
NR
4
R
5 2-tetrahydrofuran, or bicycloC 8 -C,,alkanoyl;
R
4 and R 5 being the same or different, are selected from hydrogen, Cj-C7 alkyl, phenyl (optionally substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, haloC,-C 7 alkyl, nitro, cyano and C,-C~alkoxy) or when taken together with N, form a saturated or unsaturated heterocyclic amine ring;
R'
4 and R' 5 being the same or different, are selected from C,-C 7 alkyl, cyclo(C 3
-C
8 )alkyl, phenyl (optionally substituted with I or 2 groups selected from halogen, alkyl, haloC 1 -C~alkyl, nitro, cyano and C 1
-C
7 alkoxy} or when taken together with N, form a saturated heterocyclic amine ring optionally containiing 1 or 2 additional heteroatoms selected from N, 0 or S; X is 0 or S;
R
2 and R 3 being the same or different, are selected from C 1
-C
7 alkyl, phenyl (optionally substituted with I or 2 groups selected from halogen, C 1 alkyl, haloC,-C~alkyl, nitro, cyano and C 1
-C
7 alkoxy}, C,-C 7 alkoxy, thio(C,-C 7 )alkoxy, phenoxy, thiophenoxy,
-NRR
8 (where R 7 and being the same or different, are selected from H, C,-C 7 alkyl or taken together with N, form a saturated heterocyclic ringy), or taken together with P form a 4- to 7-membered heterocyclic ring;
R
6 IS C 1 -C7 alkyl, haloC 1
-C
7 alkyl, carboC,-C 7 alkoxy, -NRgR 10 where R, and RIO, being the same or differenft, are C,-C 7 alkyl or phenyl (optionally substituted with 1 or 2 groups selected from halo, lower alkyl, haloC,-C 7 alkyl, nitro, cyano, CX- 7 alkoxy); R24 is hydrogen, halogen or Cj-C7 alkoxy;
R
25 is hydrogen or halogen; R1 4 and R1 4 b, being the same or different, are selected from hydrogen, hydroxy,
C
1
-C
6 alkyl, C 1
-C
6 alkoxy, C,-C 6 alkenyl, C 1
-C
6 alkenyl-C 1
-C
6 alkoxy, CI-C 6 alkynyl-Cl-C 6 alkoxy, CI-C 6 alkynoyloxy, poly CI-C 6 alkoxy-C 1
-C
6 alkoxy, phenyl, phenyl-C,-C 6 alkyl, tri-C 1
-C
6 alkylsilyloxy, diphenylphosphoryloxy and halogen, or RiMa and R1 4 b together form the epoxide or =CH 2 is hydrogen, C 1 -C7 alkyl, (2 2 alkoxyalkyl, C.-C 8 alkenyl, C 2 -Cs alkynyl or benzyl; the broken line between carbons 24 and 25 represents a single or double bond.
WO 93/10120 WO 9310120PCT/US92/09483 -9- Another aspect of this invention provides novel N-i marcfortine or derivatives of Formula IV:
H
3 C ,CH 3 (O)M 0 251 2 H C CH 6 24 16 N 1,2- 2NR 14 1 3 0 0 18
NR
1 8a wherein: mn is 0 or I (preferably 0); R, is selected from the group consisting of:
C
2 -C7 alkanoyl substituted with carboxy (-COOH), C,-C 7 alkanoyl, carboC,-C 7 alkoxy (-C(O)0C 1
-C
7 alkyl), -NR 4 Rs, arninocarbonyl
(-C(Q)NR
4
R
5 cyclo C 3 -Calkanoyl optionally substituted with carboxy, CI-C 7 alkanoyl, carboC,-C 7 alkoxy, -NR 4
R
5 aminocarbonyl; alkanoyloxymethylene (-CH 2
OC(O)-C,-C
7 alkyl); benzoyloxymethlene (-CFI 2 OC(O)phenyl) substituted with I or 2 groups selected from halogen, Cj-C 4 alkyl, haloC,-C 7 alkyl, nitro, cyano and C,-C~alkoxy; phenoxycarbonyl substituted with I or 2 groups selected from halogen,
C,-C
4 alkyl, haloC,-C 7 alkyl, nitro, cyano and Cl-Cqalkoxy; Mt -C(O)NR' 4
R'
5 2
)(R
3
-SR
6
C
10
-C
24 allcanoyl (-C(O)C 10 -C24alkyl);
C
1 -CAalkenoyI (-C(O)C 9
-C
23 alkenyI); or 2-tetrahydrofuran;
R
4 and R 5 being the same or different, are selected from hydrogen, C,-C7 alkyl, phenyl (optionally substituted with 1 or 2 groups selected from halogen, CI-C 4 alkyl, haloC,-C 7 alkyl, nitro, cyano and C,-C 7 alkoxy] or when taken together with N, form a saturated WO 93/10120 WO 9310120PCT/US921 09483 or unsaturated heterocyclic amine ring;
T.Z
4 and R's, being the same or different, are selected from C,-C 7 alkyl, CYClo(C 3
-C
8 )alkyl, phenyl (optionally substituted with I or 2 groups selected from halogen, alkyl, haloC,-C 7 alkyl, nitro, cyano and C,-C~alkoxy} or when taken together with N, form a saturated heterocyclic amine ring optionally containing 1 or 2 additional heteroatoms selected from N, 0 or S; X is 0 or S;
R
2 and being the same or different, are selected from C 1
-C
7 alkyl, phenyl (optionally substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, haloC,-C 7 alkyl, nitro, cyano and Cj-C 7 alkoxy), Cj-C, alkoxy, thio(C 1
-C
7 )alkoxy, phenoxy, thiophenoxy,
-NR
1 R, (where R 7 and being the same or different, are selected from H, C,-C7 alkyl or taken together with N, form a saturated heterocyclic ring), or taken together with P form a 4- to 7-membered heterocyclic ring; RO s C 1 -C7 alkyl, haloC,-Clalkyl, carboC 2 -Clalkoxy, -NR 9
R,
0 where P ad R 10 being the same or different, are C 1
-C
7 alkyl or phenyl (optionally substituted with I or 2 groups selected from halo, lower alkyl, haloCI-Clalkyl, nitro, cyano, C,-C 7 alkoxy);
R
2 is hydrogen, halogen or C,-C7 alkoxy;
R.
5 is hydrogen or halogen; is hydrogen, Cl-Cl alkyl, C 2
-C
8 alkoxyalkyl, (2 2
-C
8 alkenyl, Cl-C 8 alkynyl or benzyl; the broken line between carbons 24 and 25 represents a single or double bond.
Another aspect of this invention provides novel N-i paraherquamide or derivatives of Formula V: 0 )m 0 25 R 2 166 2 4 \19 11 10 0 Ri4a RIOb 18
N\
0 18a wherein: m is 0 or 1 (preferably 0); R, is selected from the group consisting of: WO 93/10120 WO 9310120PCT/US92/09483 -11-
C
2 -C7 alkanoyl substituted with carboxy (-COOH), Cl-C? alkanoyl, carboC 1 -C~alkoxy (-C(O)0CI-C~alkyl), -NR 4
R
5 aminocarbonyl
(-C(O)NR
4
R
5 cyclo C 3 -Cgalkanoyl optionally substituted with carboxy, CI-C 7 alkanoyl, carboCI-C 7 alkoxy, -NR 4
R
5 aminocarbonyl; alkanoyloxymethylene (-CH 2 OC(O)-C2-C~alkyl); benzoyloxymethiene (-CH 2 OC(O)phenyl) substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, haloCj-Calkyl, nitro, cyano and C,-Calkoxy; phenoxycarbonyl substituted with I or 2 groups selected from halogen,
C,-C
4 alkyl, haloC,-C 7 aLkyl, nitro, cyano and C,-CQalkoxy; Mf -C(O)NR' 4
R'
5 2
)(R
3
-SR
6 Wi C 10
-C
2 4 alkanoyl (-C(OCWC 24 alkyl);
C[-C-
4 alkenoyl (-C(O)Cq-CSalkenyl); or 2-tetrahydrofuran;
R
4 and being the same or different, are selected from hydrogen, C,-C 7 alkyl, phenyl (optionally substituted with 1 or 2 groups selected from halogen, Cj-C 4 alkyl, haloC,-C~alkyl, nitro, cyano and C,-Calkoxy) or when taken together with N, form a saturated or unsaturated heterocyclic amine ring;
R'
4 and R' 5 being the same or different, are selected from CI-C 7 alkyl, cyclo(C?-Cg)alkyl, phenyl (optionally substituted with 1 or 2 groups selected from halogen,
C,-C
4 alkyl, haloC 1 -C~alkyl, nitro, cyano and C,-C~alkoxy) or when taken together with N, form a saturated heterocyclic amine ring optionally containing I or 2 additional heteroatoms selected from N, 0 or S; X is 0Qor S;
R
2 and R 3 being the same or different, are selected from CI-C 7 alkyl, phenyl [optionally substituted with 1 or 2 groups selected from halogen, C,-C 4 alkyl, haloCI-C 7 alklcy, nitro, cyano and Cl-qatkoxyl, Cl-C? alkoxy, thio(C,-C?)alkoxy, phenoxy, thiophenoxy,
-NR
7
R
8 (where R 7 and R8, being the same or different, are selected from H, C 1
-C
7 alkyl or taken together with N, form a saturated heterocyclic ring), or taken together with P form a 4- to 7-membered heterocyclic ring;
R
6 is alkyl, haloC,-C~alkyl, carboC 1 -Ca~koxy, -NR 9
R
10 where R 9 and RI., being the same or different, are C,-C7 alkyl or phenyl (optionally substituted with 1 or 2 groups selected from halo, lower alkyl, ha~oC 1 -C~alkyl, nitro, cyano, C 1
-C
7 alkoxy); WO 93/10120 WO 9310120PCr/US92/09483 -12-
R
24 is hydrogen, halogen or CI-C 7 alkoxy;
R
25 is hydrogen or halogen;
R,
4 and R1 4 b, being the same or different, are selected from hydrogen, hydroxy, C 1
-C
6 alkyl, C,-C 6 alkoxy, C,-C 6 alkenyl, C,-C 6 alkenyl-CI-q6 alkoxy, C,-C 6 alkynyl-C 1
-C
6 alkoxy,
C,-C
6 alkynoyloxy, poly C,-C 6 alkoxy-C,-C 6 alkoxy, phenyl, phenyl-C,-C 6 alkyl, tri-C 1
-C
6 alkylsilyloxy, diphenylphosphoryloxy and halogen, or R1 4 and R1 4 b together form the epoxide or is hydrogen, CI-C 1 alkyl, C 2 alkoxyalkyl, C,-CB alkenyl, Cq-C 8 alkynyl or benzyl; the broken line between carbons 24 and 25 reprdsents a single or double bond, The carbon content of various hydrocarbon containing moieties is indicated by a prefix designating the minimum and maximum numnber of carbon atoms in the moiety, the prefix
C
1 indicates a carbon atoms content of the integer to the integer carbon atoms, inclusive. Thus, C,-C 3 alkyl refers to alkyl of 1-3 carbon atoms, inclusive, or methyl, ethyl, propyl, and isopropyl.
With resp~t to the above, "C 1
-C
7 alkyl" is intended to include those alkyl groups of from I to 7 carbon atoms in either a straight or branched represents a double bond between chain. Examples of such lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, secbuty), tert-butyl, pentyl, hexyl, heptyl and the like.
Cyclo(C.
3
-C
8 )alkyl is intended to include alkyl rings of 3 to 8 members. Examples of cyclo(q3-C,)alkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicycloll2.2. 1]heptyl, and the like.
alkoxy is intended to include those alkoxy groups of from I to 8 carbon atoms in either a straight or branched chain. Examples of such C,-C 8 alkoxy groups are methoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, pentoxy, hexoxy, heptoxy, and the like.
C
2
-C
7 alkanoyl is intended to include those alkanoyl groups of from 2 to 7 carbon atoms in either a straight or branched chain. Examples of such C 2
-C
7 alkanoyl groups include acetyl, propionyl, isopropionyl, bDatyryl, pentanoyl, hexanoyl and the like.
C
10 -C24alkanoyl (-C(O)C 9
-C
23 alkyl) is intended to include those alkanoyl groups of from to 24 carbon atoms in either a straight or branched chain. Examples of such C 10
C
2 q 4 alkanoyl groups include decanoyl [-C(O)(CH 2 9
CH
3 1, lauroyl [-C(O)(CH)I 0
CH
3 1, tridecanoyl
(CHA)
1
CH
3 myristoyl 12
CH
3 pentadecanoyl [-C(Q)(C1 2 1 3
CH
3 1 palmitoyl [-C(O)(CH 2 14
CH
3 1, magaroyl [-C(O)(C1 2 )j 5
CH
3 stearoyl
[-C(Q)(CH
2 16
CH
3 arachidoyl [-C(Q)(CH 2 8
CH
3 1, heneicoisanoyl [-C(O)(C1 219
CH
3 1, behenoyl K-(Q)(CH 2 20
CH
3 tricosanoyl [-C(O)(CH 2 1
CH
3 tetracosanoyl
[-C(O)(CH
2 22 and the like.
C
10
-C
24 alkenoyl (-C(Q)C 9
-C
23 alkenyl) is intended to include those unsaturated groups of WO 93/10120 PCT/U592/09483 -13from 10 to 24 carbon atoms in either a straight or branched chain. Examples of such Co-C, alkenoyl groups include undecylenoyl [-C(O)(CH 2
),CH:CHCH
3 oleoyl
CH:CH(CH
2 7
CH
3 linoloyl 7
CH:CH.CH
2
.CH:CH(CH
2 4 C1] 3 and the like.
The term alkoxyalkyl" is intended to include those lower alkoxy substituted lower alkyl groups containing from 2 to 8 carbon atoms and from 1 to 3 oxygen atoms in either a straight or branched chain. Examples of such C(-C alkoxyalkyl groups include methoxymethyl, methoxyethoxymethyl, methoxyethoxyethoxymethyl, ethoxyethyl, and the like. Examples of alkoxymethyl are methoxymethyl, ethoxymethyl, propoxymethyl, butoxymethyl, pentoxymethyl, butoxymethyl, pentoxymethyl, hexoxymethyl, and heptoxymethyl, and isomeric forms thereof.
The term alkanoyloxymethylene is intended to include those alkanoyloxy substituted methylenes containing from 2 to 8 carbon atoms in either a straight or branched chain.
Examples of such C 2 -C alkanoyloxymethylene groups include acetoxymethyl, tert-butoxymethyl, n-propoxymethyl, valeroxymethyl and the like.
The term "substituted benzoyloxymethylene" is intended to include those benzoyloxymethyl groups in which the benzene ring is substituted with from 0 to 3 substituents selected from lower alkyl, trifluoromethyl, C 1
-C
7 alkoxy, nitro, or cyano groups, and halogen atoms.
The term "Isubstituted benzenesulfonyl" is intended to include those benzenesulfonyl groups in which the benzene ring is substituted with from 0 to 3 substituents selected from lower alkyl, trifluoromethyl, CI-C, alkoxy, nitro, or cyano groups, and halogen atoms.
The term "halogen" is intended to include mie halogen atoms fluorine, chlorine, bromine and iodine.
The term "halo C,-Calkyl" is intended to include those halogen substituted C 1 alkyl groups containing from I to 7 carbon atoms in either a straight or branched chain and from 1 to 3 halogen atoms. Examples of haloC-Calkyl include fluoromethyl, 2-bromoethyl, 3-chloropropyl, 5-iodopentyl, trifluoromethyl, and the like.
The term alkenyl" is intended to include those lower alkyl groups containing from 2 to 8 carbon atoms in either a straight or branched chain which contains 1 to 2 carbon double bonds. Examples of such C 2 -C alkenyl groups include allyl, 3-butenyl, 2,4-pentadienyl, hexenyl, and the like.
The term alkynyl" is intended to include those alkynyl groups containing from I to 8 carbon atoms in either a straight or branched chain which contains 1 to 2 carbon-carbon triple bonds. Examples of such C-C alkynyl groups include propargyl, 2-butynyl, 2,4-pentadiynyl, 5-hexynyl, and the like.
Examples of "alkoxycarbonyl" 2 7 alkoxy) include ethoxycarbonyl, WO 93/1012G WO 931012GPCr/US92/09483 -14isopropoxycarbonyl, methoxycarbonyl, butoxycarbonyl, hexoxycarbonyl and the like.
C,-C
7 alkanoyl is intended to include alkyl groups of from I to 7 carbon atoms in either a straight or branched chain. Examples of C,-C 7 alkanoyl include acetyl, pmopionyl, iso-butyryl, valeryl, 5-methyihexanoyl, and the like.
Examples of aminocarbonyl -C(=O)NR 4
R
5 include dimethylaminocarbonyl, propylmethylaminocarbonyl, dibutylaminocarbonyl, isopropylaminocarbonyl, hexylaminocarbonyl and the lie.
Examples of aminothiocarbonyl -C(=S)NR 4
R
5 include dimethylaminothiocarbonyl, propylmethylaminothiocarbonyl, dibutylaminothiocarbonyl, isopentylarniinothiocarbonyl, bexylaminothiocarbonyl and the like.
Examples of the group 3 include diethyl thiophosphoryl, phenylmethoxyphosphonyl, 2-thioxo-1I,3,2-dioxaphosphorinanyl, N,Ndimethylmethoxyphosphoramnidyl, diphenyiphosphinyl and the like.
Examples of -SR 6 include 2,4-dinitrobenzenesulfenyl, dimethylatninosulfenyl, ethoxycarbonylsulfenyl, trichioromethylsulfenyl, 4-morpholinosulfenyl and the like.
Examples Of -SO 2
NR
4 Rs include dimethylsulfamoyl, phenylmethylsulfamoyl, 4-morpholinosulfamoyl, piperidinylsulfamoyl and the like.
The term "P containing heterocyclic ring" is intended to include 1,3-dioxa-2phosphorinane, 1 -aza-3-oxa-2-phospholane, I ,3-diaza-2-phospholane, 1-thia-3-oxa-2phospholane, and the like.
Examples of heterocyclic amine rings according to -NR 4
R
5
-NR'
4
R'
5 and are: 4-morpholine, 4-phenyl-l1-piperazine, 4-(2-pyridinyl)-lI-piperazine, 2,6-dimethyl-4-morpholine, 1-pyrrolidine, 4-methyl-I -piperazine, I -piperidine, 4-phenyl- 1 -piperidine thiazolidine, 4-phenyl- 1 ,2,3,6-tetrahydropyridine, 4-phenylpiperidine, ethyl prolinate, tetrahydrofurylamine, 3-pyrroline, thiazolidine-4-carboxylic acid, WO 93/10120 WO 9310120PCT/US92/09483 thiomorp-holine, nipecotamide, 2-methylpiperidine, 3-methylpiperidine, 4-methylpiperidine, N-methylpiperazine, I -methyihomopiperazine, I -acetylpiperazine, N-carboethoxypiperazine, Pharmaceutically acceptable salts means salts useful for administering the compounds of this invention -and include mesylate, hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, acetate, propionate, lactate, ma'eate, malate, succinate, tartrate, and tht-, like. These salts may be in hydrated form.
Preferred compounds of this invention are compounds of Formula I (as well as Formula 11, 111, IV ar V) where R 24 and R 25 are :alydrogen; is hydrogen, C,-C 4 alkyl, C 2
-C
4 alkoxyalkyl, q 2
-C-
4 alkenyl or benzyl; and the broken line between carbons 24 and 25 represents a double bond.
Examples of the compounds of Formula 11 of th-is invention are as follows: I 8-thiomarcfortine A I-acetoxymethyl-18-thiomarcfortine A I -diethoxyphosphoryl- 18-thiomarcfortine A I -dimethylsulfamr~oyl- 18-thiomarcfortine A I -cyclopropylcarbonyl-1I8-thiomarcfortine A 2-bicyclo 12.2.1 ]heptanoyl-1 8-thiomarefortine A 1-(l-piperidinyl)thiocarbonyl-1 8-thiomarcforiine A I -succinoyl-1 8-thiomartfortine A I -(4-morpholinosulfenyjl)-1 8-thiomarcfortine A 1 -(2,4-dinitrobenzenesulfenyl)-1I8-thiomarcfortine A 24-propoxy-24,25-dihydro- 18-thiomarefortine A 1 -(p-toluenesulfonyi)-1I8-thiomarcfortinie A 1-acetyl-1 8-thioruarcfortine A 1-methyl-I 8-thiomarcforfine A I -benzyl- 18-thiomarcfortine A 1-dimethylcarbamnoyl- 18-thiomarcfortine A 1-methoxycarbonyl-18-thiomarcfortine A 24,25-dihydro- 18-thiomarcfortine B WO 93/10120 WO 93/1120 PCMUS92/O9483 -16- 24-rnethoxy-24,25-dihydro-I 8-thiomarcfortine B I -(p-toluenesulfonyl)-1I8-thiomarcfoitine B 1 -ethyl-I 8-thiomarcfortine B I-benzyl-1I8-thiorarcfortine B 18a-ethyl-18-thiomarcfortine B 1 8a-benzyl- 18-thiomarcfortine B I 8a-methoxyethoxym ethyl- 1 8-thiomarcfortine B 18a-aIlyl-18-tbiomarcfortine B I 8a-propargyl-1I8-thiomarcfortine B 1 8a-ethyl-24-methioxy-24,25-dihydro- 18-thiomarcfortine B 1, 1 8a-b is-ethyl- I 8-thiomarcfortine B 1,1 8a-bis-benzyl- 18-thiomarcfortine B 1 8a-ethyl-24-.methoxy-1I8-thiomarcfortine B 1 -(2,4-dinitrobenzene sulfonyl)- I a-ethyl-24-methoxy-24,25-dihydro-1I8-thiomarcfortine B 18a-ethyl-24,25 dihydro-18-'.hiomarcfortine B 24,25 dihydro-I 8-thiomarcfortine C I -(p-bromobenzene sulfonyl)- I -tliomarcfortine C I -propionyl-1I8-thiomarcfortine C 1 -propyl- 18-thiomarefortine C 1-benzyl-18-thiomarcfortine C 1 8a-propyl- 18-thiomnarcfortine C 1 8a-benzyl- I8-thiomarcfortine C I 8a-rnethoxyethoxyinethyl -1 8-thiomarcfortine C 18-alyl-18-thiomarcfortine C I 8a-propargyl-1 8-thiomarcfortine C 1,1 8a-bis-propyl- 18-thiomarcfortine C 1, 1 8a-bis-benzyl- I 8-thiomarcfortine C I 8-thiomarcfortine C 1 -(4-carbethoxy-4 ,3-thiazolidinin-3-yl)carbonyl- 18-thiiomarcfortine A 1-palmitoyl18-thiomarcifortine A I -(4-morpholinocarbonyl)- 18-thiomarcfortine A.
Examiples of th'e compounds of Formula III of this invention are as follows: I 8-thioparaherquamide 24,25-dihydro-I 8-thioparaberquamide 14-O-methyl- 18, thioparaherquamide 14-0-ethyl- 18-thioparaherquamide WO 93/10120 WO 93/10120PCr/US92/09483 -17- 14-0-butyl- 1 8-thioparaherquamide 14-0-bepzyl- 18-thioparaherquamnide 14-0-ally-i 8-thioparaherquaxnide 14- 0-propargyl- 1 8-thioparaherquaxnide 1 4-0-methoxymethyl- 1 8-thioparaherquarnide 14-0-methoxy-ethoxy-I 8-thloparaherquamide 14-0-methoxy-ethyoxy-methyl- 1 8-thioparaherquaniide 17-methyl- I 8-thioparaherquamnide 17-methylene- 1 8-thioparaherquamide 1 -N-(p-toluenesulfonyl)- I 8-thioparaherquamide 24-methoxy-24,25-dihydro- 1 8-thioparaherquamide Example., of the compounds of Formula IV of this invention are as follows: 1-diethoxythiophosphoryl-marcfortine A 1 -phenylmethoxyphosphonyl-marcfortine A l-cyclohexylcarhonyl-marcfortine A 1 -piperidLiyl)thiocarbonyl-mnarcfortine A I -N-(4-carbethoxy- 1,3-thizolidinin-3-yl)carbonyl-marcfortine A 1-succinoyl-marcfortine A 1-(4-exopentanoyl)-marcfortine A 1-dimethylaininoaretyl-marcfortine A 1-N,N-dimethylsuceiamidoyl-marcfortine A 1-cyclopropylcarbonyl- 18a-ethyl-marcfortine B 1 -dimethylamninosulfenyl- 18a-N-benizyl-marcfortine B 1-ethoxycarbonylsulfenyl- 18a-N-methoxyethoxymethyl-marcfortine B 1 -diphenylphosphinyl-1I8a-N-allyl-marcfortine B I -(3-acetoxy)propionyl- 18a-N-propargyl-marcfortine B 1 -(2,4-dichlorophenoxy)carbonyl-1I8a-N-ethyl-24-methoxy-24,25-dihydro-marcfortine- B 1-N-(2,4-dinitrobenzenesulfenyl)-1I8a-N-ethyl-niarcfortine B 1-N-(p-bromobenzenesulfenyl)-marcfortine C 1-acetoxymethy' mnarcfortine C 1-diethoxythiophosphoryl-marcfortine C 1 -phenylmethoxyphosphonyl-marcfortine C 1-cyclopropylcarbonyl-marcfortine C 1 -cyclohexylcarbonyl-marefortine C 1 -piperidinyl)thiocarbonyl-marcfor-ine C 1 -N-(4-carbethoxy- 1,3-thiazolidinin-3-yl)carbonyl-marcfortine C WO 93/10120 W093/0120PCr/US92/09483 1-succinoyl-marcfortine C 1 -(4-morpholinosulfenyl)-marcfortine C 1 -phenoxycarbonyl-marcfortine C I -(4-oxopentanoyl)-marcfortine C 1-dimethylaminoacetyl-marcfortine C 1 -N,N-dimethylsuccinamidoyl-marcfortine C 1-palmitoyl-marcfortine A; preferably I -cyclopropylcarbonyl-marcfortine A I -phenoxycarbonyl-marcfortine A 1 -palm itoyl-marcfortine A 1 [(4-ntrophenyl)oxy]carbonyl]-marcfortine A 1 -(1.piperidinecarbonyl).rnarcfortine A 1- [[4-(ethoxycarbonyl)piperazin- 1-yl]carbonyl]-marcfortine A 1-[[14-(benzyl)piperazin- 1-yl]carbonyl]-marcfortine A 1- 1,3-benzodioxol-5-yl-rnethyl)piperazin- 1-y1}carbonyl]-marcfortine A 1 [4-(methyl)piperazin- 1-yl]carbonyl]-marcforiine A 1 ,3-dihydro- 1,3-dioxo-2H-isoindol-2-yl)-marcfortine A 1 -I[4-(pyridin-2-yl)piperazin- 1-yI]carbonyl]-marcfortine A 1-[[4-(phenyl)piperazin-1-yl]carbony1]-marcfortine A 14 14-(chlorocarbonyl)piperazin- 1-yl]carbonyll-marcfortine A -(phenyl)piperidin- 1-yIlcarbonyl]-marcfortinie A 1 [4-(dimethyl)piperidin- 1-ylllcarbonyl]-marcfortine A 14 [4-(5-chloropyridazin-3-yl)piperazin- 1-yllcarbonyl]-marcfortine A 1 [4-(4-,,hlorophenyl)pirprazin- 1-yl]carbonyl]-rnarcfortine A 1-[[4-(trichloroethoxycarbonyl)piperazin- 1-yllcarbonyl]-marcfortine A 14 [4-(3-trifluoromethyl-thiadiazol-1-yl)piperazin-1-yl]carbonyl]-rnarcfortine A 1-acetoxymethyi-marcfordne A 1 .',,-c~nitobenzenesulfeny1)-marcfortine A 1 -(4-mairtholinosulfeny1)-rnarcfortine A 1 -(trichloromethylsulfenyl)-marcfortine A 1-(methoxycarbonylsulfenyl)-r-narcfortine A 1 -(benzenesulfenyl)-marcfortine A 1-(2-tetrahydrofuranyl)-marcfortine A, and more preferably, I -(4-morpholinocarbonyl)-marcfortine A or 1-(4-morpholinocarbonyl)-marcfortine A N-oxide.
Examples of the com-npounds of Formula V of ths invention are as follows: WO 93/10120 WO 9310120PCT/US92/09483 -19- I -acetoxymethyl-paraherquamide 1-diethoxythiophosphor-yl-paraherquaxnide I -pbenylmethoxyphosphonyl-paraherquamide 1 -cyclopropylcarbonyl-paraherquarnide 1 -cyclohexylcarbonyl-paraherquaniide I 1-piped dinyl)thiocarbonyl-paraherquamide l-N-(4-carbethoxy- I ,3-thiazolidinin-3-yl)carbonyl-paralierquamide 1 -succinoyl-paraherquamide 1 -(4-morpholinosulfenyl)-paraherquamide 1 -phenoxycarbonyl-paraherquamnide I -(4-oxopentanoyl)-paraherquamide I -dimethylaminoacetyl-paraherquamide I -N.N-dimethylsuccinamidoyl-parahierquamide I -palmnitoyl-paraherquamnide 1 -(4-morpholinocarbonyl)-paraherquamide.
The compounds of Formula II are prepared by the following procedures: It has been found, unexpectedly, ijiat treatment of the Marcfortines A, B, and C, or suitably substituted C-24, C-25, N-i and N-18a derivatives thereof, with phosphorous pentasulfide or, preferably, 2,4-bis(methoxyphenyl)-1 ,3-dithia-2,4-diphosphetane-2,4-disulfide (Lawesson's reagent) selectively provides the corresponding 18-thio- derivatives. Suitable C-24, C-25, N-I and N-1I8a substituted marcfortirne A, B and C derivatives for this reaction are readily prepared by the procedures given in U.S. paten, 4,923,867, th, disclosure of which is incorporated herein by reference. The reactions are conducted in a suitable inert solvent such as pyridine, collidine, toluene (preferred), xylene, dioxane, tetrahydrofuran, and the like, at temperatures from 100 to 1800 preferably 800 to 1400.
Alternatively, the C-24, C-25 and N-1 derivatives can be prepared from the 18-thiomarcfortines. For example, a large series of 18-thiomarcfortine analogs can be prepared by alkylation or acylation of N-i of the N-18a substituted 18-thiomarcfortines. These derivatives may be easily prepared by sequential treatment of a solution of 18-thiomarcfortine A, N-i 8a-substituted 1 8-thiomarefortine B, or N-i 8a-substituted 1 8-thiomarcfortine C in an aprotic organic solvent such as tetrahydrofuran, ether, benzene and the like with an excess of a strong base such as potassium hydride (preferred), sodium hydride, butyllithium, potassium tertbutoxide, and the like followed by a suitable alkylating or acylating agent at temperatures ranging from 0 0 C to 50 0 C for 0.25 to 48 hours. Suitable alkylating agents include alkyl bromides, alkyl iodides, alkyl sulfonates, alkenyl iodides, aikynyl bromides, alkoxyalkyl chlorides, and the like. Suitable acylating agents include acyl anhydrides, acyl chlorides, acyl WO 93/10120 PCT/US92/09483 bromides, substituted benzenesulfonyl chlorides, substituted benzenesulfonic anhydrides, sulfenyl chlorides, isocyanates, carbamoyl chlorides, chloroformates, and the like.
An additional series of derivatives can be generated by modification of the C24-C25 double bond of 18-thiomarcfortines A, B and C. The 24,25 dihydro analogs are readily prepared by stirring a solution of the appropriate 18-thiomarcfortine in an alcoholic solvent such as methanol, ethanol, propanol and the like with a catalyst such as palladium, platinum, tris (triphenylphosphine)-chlororhodium and the like in the presence of hydrogen gas. The product, which is a 24,25-dihydro-18-thiomarcfortine analog, can be isolated and purified by using techniques known to those skilled in the art. Note that the reactions described above for modification of other portions of the 18-thiomarcfortine structure may also be applied to 24,25-dihydro 18-thiomarcfortine analogs to prepare the corresponding 24,25-dihydro analogs.
Additional C24-C25 double bond modified analogs of the 18-thiomarcfortines may be prepared via the 24,25 dibromide which is easily prepared by treating a solution of a 18-thiomarcfortine in a halogenated solvent such as dichloromethane, chloroform, carbon tetrachloride and the like with 1 molar equivalent of bromine at temperatures ranging from -20 0 C to 25 0 C for 0.25 to 8 hours. This process affords the corresponding 24,25-dibromo 24,25-dihydro-18-thiomarcfortine derivative which can be isolated and purified by using techniques known to those skilled in the art. Note that the 24,25 dichloro analog may be prepared by substituting chlorine for bromine in the process described above. The 24,25-dibromo 24,25-dihydro-18-thiomarcfortine analogs described above are useful intermediates for the preparation of additional derivatives. Thus, treatment of a solution of the dibromide in an alcoholic solvent such as methanol, ethanol, propanol, and the like with a strong base such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) at temperatures ranging from OC to 30°C for 0.25 to 24 hours affords 24-alkoxy-25 bromo-24,25 dihydro-18-thiomarcfortine analogs which can be isolated and purified by using techniques known to thtse skilled in the art. These 24 alkoxy, 25-bromo derivatives can be debrominated by treatment of a solution of the compound in an aprotic organic solvent such as benzei,-; toluene, hexane, and the like with a tin hydride reducing agent such as tri-butyl tin hydride, tri-phenyl tin hydride and the like with or without the addition of a radical initiator such as azobis-isobutyronitrile (AIBN) at temperatures ranging from 25 0 C to 120 0 C for 0.5 to 48 hours.
This process affords the corresponding 24-alkoxy-18-thiomarcfortine derivatives (R lower alkoxy in the general structure) which can be isolated and purified by using techniques known to those skilled in the art.
General procedures for the preparation of heteroaromatic N-oxides can be found in Chapter II of "Chemistry of the Heterocyclic N-Oxides", A.R. Katritzky and J.M. Lagowski, published 1971 Academic Press (Vol. 19 of ORGANIC CHEMISTRY A Series of Monographs). Typically the N-oxide is formed by reaction with a percarboxylic acid in an WO 93/10120 PCT/US92/09483 -21appropriate solvent. Most suitably an aromatic peracid in a non-polar solvent is used, since the reaction may usually be carried out at room temperature. Suitable aromatic peracids include perbenzoic acid, chloroperbenzoic acid and perphthalic acid.
The compounds of Formula III are prepared by the following procedures: Treatment of paraherquamide and derivatives thereof, with phosphorous pentasulfide or, preferably, 2,4-bis(methoxyphenyl)-l,3-dithia-2,4-diphosphetane-2,4-disulfide (Lawesson's reagent) selectively provides the corresponding 18-thio- derivatives. Suitable paraherquamidle derivatives for this reaction are readily prepared by the procedures given in U.S. patent 4,978,656 and 4,873,247 as well as WO 91/09961, the disclosures of which are incorporated herein by reference. The reactions are conducted in a suitable inert solvent such as pyridine, collidine, toluene (preferred), xylene, dioxane, tetrahydrofuran, and the like, at temperatures from 100 to 1800 preferably 800 to 1400.
Alternatively, the C-24, C-25 and N-I derivatives can be prepared from the 18-thioparaherquamide. For example, a large series of 18-thioparaherquamide analogs can be prepared by alkylation oi acylation of N-l of the N-18a substituted 18-thioparaherquamide.
These derivatives may be easily prepared by sequential treatment of a solution of 18-thioparaherquamide in an aprotic organic solvent such as tetrahydrofuran, ether, benzene and the like with an excess of a strong base such as potassium hydride (preferred), sodium hydride, butyllithium, potassium tert-butoxide, and the like followed by a suitable aikylating or acylating agent at temperatures ranging from 0 C to 50 0 C for 0.25 to 48 hours. Suitable alkylating agents include alkyl bromides, alkyl iodides, alkyl sulfonates, alkenyl iodides, alkynyl bromides, alkoxyalkyl chlorides, and the like. Suitable acylating agents include acyl anhydrides, acyl chlorides, acyl bromides, substituted benzenesulfonyl chlorides, substituted benzenesulfonic anhydrides, sulfenyl chlorides, isocyanates, carbamoyl chlorides, chloroformates, and the like.
An additional series of derivatives can be generated by modification of the C24-C25 double bond of 18-thioparaherquamides. The 24,25 dihydro analogs are readily prepared by stirring a solution of the appropriate 18-thioparaherquamide in an alcoholic solvent such as methanol, ethanol, propanol and the like with a catJyst such as palladium, platinum, tris (triphenylphosphine)-chlororhodum and the li&c iii the presence of hydrogen gas. The product, which is a 24,25-dihydro-18-thioparaherquamide analog, can be isolated and purified by using techniques known to those skilled in the art. Note that the reactions described above for modification of other portions of the 18-thioparaherquamide structure may also be applied to 24,25-dihydro 18-thioparaherquamide analogs to prepare the corresponding 24,25-dihydro analogs. Additional C24-C25 double bond modified analogs of the 18-thioparaherquamides may be prepared via the 24,25 dibromide which is easily prepared by treating a solution of a 18-thioparaherquamide in a halogenated solvent su"h as dichloromethane, chloroform, carbon WO 93/10120 PCIYUS92/09487 -22tetrachloride and the like with 1 molar equivalent of bromine at temperatures ranging from 0 C to 25 0 C for 0.25 to 8 hours. This process affords the corresponding 24,25-dibromo 24,25-dihydro-18-thioparaherquamide derivative which can be isolated and purified by using techniques known to those skilled in the art. Note that the 24,25 dichloro analog may be prepared by substituting chlorine for bromine in the process described above. The 24,25-dibromo 24,25-dihydro-18-thioparaherquamide analogs described above are useful intermediates for the preparation of additional derivatives. Thus, treatment of a solution of the dibromide in an alcoholic solvent such as methanol, ethanol, propanol, and the like with a strong base such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) at temperatures ranging from 0°C to 30 0 C for 0.25 to 24 hours affords 24-alkoxy-25 bromo-24,25 dihydro-18-thioparaherquamide analogs which c'i be isolated and purified by using techniques known to those skilled in the art.
These 24 alkoxy, 25-bromo derivatives car. be debrominated by'treatment of a solution of the compound in an aprotic organic solvent such as benzene, toluene, hexane, and the like with a tin hydride reducing agent such as tri-butyl tin hydride, tri-phenyl tin hydride and the like with or without the addition of a radical iniator such as azobis-isobutyronitrile (AIBN) at temperatures ranging from 25 0 C to 120 0 C for 0.5 to 48 hours. This process affords the corresponding 24alkoxy-18-thioparaherquamide derivatives lower alkoxy in the general structure) which can be isolated and purified by using techniques known to those skilled in the art.
General procedures for the preparation of heteroaromatic N-o-ides can be found in Chapter II of "Chemistry.of the Heterocyclic N-Oxides", A.R. Katritzky and J.M. Lagowski, published 1971 Academic Press (Vol. 19 of ORGANIC CHEMISTRY A Series of Monographs). Typically the N-oxide is formed by reaction with a percarboxylic acid in an appropriate solvent. Most suitably an aromatic peracid in a non-polar solvent is used, since the reaction may usually be carried out at room temperature. Suitable aromatic peracids include perbenzoic acid, chloroperbenzoic acid and perphthalic acid.
The compounds of Formula IV are prepared by alkylation or acylation of N-1 of marcfortine A and N-18a substituted marcfortines B and C. Th'se derivatives may be easily prepared by sequential treatment of a solution of marcfortine A, or N-18a substituted marcfortines B or C in an aprotic organic solvent such as tetrahydrofuran, ether, benzene and the like with an excess of a strong base such as potassium hydride (preferred), sodium hydride butyllithium, potassium tert-butoxide, and the like followed by a suitable alkylating or acylating agent at temperatures ranging from 0 C to 50°C for 0.25 to 48 hours. Suitable alkylating or acylating agents include alkanoyloxyalkyl bromides, aminosulfenyl chlorides, phosphoryl chlorides, phosphonyl chlorides, acyl anhydrides, acyl chlorides, acyl bromides, substituted benzenesulfenyl chlorides, carbamoyl chlorides, substituted phenoxychloroformates, and the like.
Treatment of marcfortine A and N-18a substituted marcfortines B and C and C24-C25 modified WO 93/10120 PCT/US92/09483 -23marcfortines under these conditions affords I-N-substituted analogs which can be isolated and purified by using techniques known to those skilled in the art.
The compounds of Formula V are prepared by alkylation or acylation of N-l of paraherquamide. These derivatives may be easily prepared by sequential treatment of a solution of paraherquamide in an aprotic organic solvent such as tetrahydroftran, ether, benzene and the like with an excess of a strong base such as potassium hydride (preferred), sodium hydride butyllithium, potassium tert-butoxide, and the like followed by a suitable alkylating or acylating agent at temperatures ranging from 0°C to 50°C for 0.25 to 48 hours. Suitable alkylating or acylating agents include alkanoyloxyalkyl bromides, aminosulfenyl chlorides, phosphoryl chlorides, phosronyl chlorides, acyl anhydrides, acyl chlorides, acyl bromides, substituted benzenesulfenyl chlorides, carbamoyl chlorides, substituted phenoxychloroformates, and the like.
Treatment of paraherquamide under these conditions affords 1-N-substituted analogs which can be isolated and purified by using techniques known to those skilled in the art.
General procedures for the preparation of heteroaromatic N-oxides can be found in Chapter II of "Chemistry of the Heterocyclic N-Oxides", A.R. Katritzky and J.M. Lagowrki, published 1971 Academic Press (Vol. 19 of ORGANIC CHEMISTRY A Series of Monographs). Typically the N-oxide is formed by reaction with a percarboxylic acid in an appropriate solvent. Most suitably an aromatic peracid in a non-polar solvent is used, since the reaction may usually be carried out at room temperature. Suitable aromatic peracids include perbenzoic acid, chloroperbenzoic acid and perphthalic acid.
PREPARATION OF STARTING MATERIAL Paraherquamide is isolated from Penicillium Sp. IMI 332995 and/or Penicillium charlessi MF 5123 (ATCC 20841) using standard fermentation and isolation techniques. The isolation is described in detail in US Patent 4,873,247 and 4,978,656 as well as EP 390532-A, EP-301742-A and WO 91/09961 (all of which are incorporated herein by reference).
PREPARATION OF STARTING MATERIAL N-18a substituted marcfortines B and C and C24-C25 modified marcfortines are readily prepared by procedures given in U.S. Patent 4,923,867, the disclosure of which is incorporated herein by reference.
Marcfortines A, B and C are isolated, along with the previously known roquefortine, as fungal metabolites of Penicillium roqueforti using standard fermentation and isolation techniques. The isolation, as well as the analytical and structural characteristics of marcfortine A, are described in detail in Polonsky et al Journal of the Chemical Society Chemical Communications 1980, 601-602. The isolation, as well a; the analytical and structural WO 93/10120 PCT/US92/09483 -24characteristics of marcfortines B and C, are described in detail in Polonsky et al Tetrahedron Letters 1981, 22, 1977-1980.
Alternatively, and more preferably, Marcfortines A and C may be isolated from Penicillium sp. UC7780 (strain number in Upjohn Culture Collection, UC 7780, The Upjohn Company, Kalamazoo, MI). This strain was isolated from a soil sample collected in Illinois, on \3 Se.pkq be-r lAo I deposited in the U.S. Department of Agriculture patent culture collection in Peoria, IL and given the accession number NRRL 18887. To further characterize the fungus a taxoncmy sudy was done following the methods and materials described by I. John Pitt, The Genus Penicillium, Academic Press, London, (1979). Spore and hyphae surfaces were examined by scanning electron microscopy cording to the methods of Dietz, A. and Matthews, J, Appl. Microbiology 18:694-696 (1969). int coidiophores are visualized by light microscopy Onions et al., Smith's Introduction to Industrial Mycology, John Wiley and Sons, New Yoik, pp 301-302 (1979)] after slide culture(s) are prepared: A glass petri dish containing glass beads, microscope slide, and coverslip are sterilized. A small block of potato dextrose agar is placed on the slide and inoculated on four sides with the funguE culture. The coverslip is set on the inoculated agar block and sterile water added to maintain moisture. The chamber is incubated for six days at 24 OC. A slide is prepared by removing the coverslip and placing it on a drop of lactophenol cotton blue stain.
The characteristics of Penicillium sp. UC 7780 (NRRL 18887) are as follows: Morphology a biverticillate penicillus (two branch points between conidium and stipe).
These branches (retulae) support the phialides or conidia bearing structures. Conidiophores (approximately 35 pm) terminated in verticils of 2-5 (10-14 pm) metulae. Phialides were ampullifcrm (like an ancient Greek wine jar) in verticils of 2-5 (7 pm). Conidia were smooth and spheroidal (2 pm) typically appearing in long columns. The stipe walls were smooth.
The culture was inoculated onto three petri dishes of Czapek yeast agar (CYA), one being 6 cm in diameter and one each of malt extract agar (MEA) and 25% glycerol nitrate agar Inoculation was made from a semisolid suspension (0.5 ml of 0.2% agar with 0.05% Tween 80). An inoculating loop of conidia was added to the tube and mixed. A loop of suspension was inoculated in a pattern of three sites per plate. A needle was used to stab inoculate the 6 cm plate. The incubation regime was: one CYA plate plus the MEA and plates at 24°C, one CYA plate at 37 0 C, and the 6 cm CYA plate at 5°C. After seven days the colony diameters, colors, and other characters were recorded and are set forth ii Table I. On potato dextrose agar (PDA, Difco) agar, a deep red color on the bottom or reverse of the colony is produced.
No sexual stage was noted. This results in the culture (NRRL 18887) being keyed in thr Penicillium Key to subgenera. Within the Penicillium Key, the penicillus type alone WO 93/10120 PC/US92/09483 determines the subgenus to which a species is allocated. This species has several characteristics that distinguish it from the Biverticillium subgenus even though its penicillus is biverticilliate.
The species consistently produces colonies greater than 10 mm in diameter in 7 days on glycerol nitrate agar. The metulae appear longer than the phialides and are in verticils of 2-5. These characteristics place this Penicillium sp. (NRRL 18887) into the subgenus Furcatum.
The foregoing description is illustrative of a strain of Penicillium sp. UC 7780 (NRRL 18887) which can be employed in the production of Marcfortine and derivatives thereof.
However, the present invention also embraces mutants of the above described microorganism.
For example, those mutants which are obtained by natural selection or those produced by mutating agents including ionizing radiation such as ultraviolet irradiation, or chemical mutagens such as nitrosoguanidine or the like treatments are also included within the ambit of this invention.
This description is also desired and intended to include inter- and intraspecific genetic recombinants produced by genetic techniques well known to those skilled in the art, such as. for example, conjugation, transultion and genetic engineering techniques.
Penicillium sp. UC 7780 (NRRL 18887) may be cultivated under aerobic condition in the same manner as commonly employed in the art for cultivation of a known strain of the genus Penicillium.
As medium components may be employed any of the well-known nutrient materials for Penicillium. For instance, as an assimilable carbon source, glucose, glycerol, maltose, dextrin, starch, lactose, sucrose, molasses, soybean oil, cotton seed oil, etc., preferably glucose and glycerol may be employed and, as an assimilable nitrogen source, soybean meal, peanut meal, cotton seed meal, fish meal, com steep liquor, peptone, rice, bran, meat extract, yeast, yeast extract, sodium nitrate, ammonium nitrate, ammonium sulfate, etc. may be used. And, such inorganic salts as sodium chloride, phosphates, calcium carbonate, etc. may be added to a culture medium. A minor amount of a metal salt may also be added, if necessary. Further, a minor amount of a heavy metal may be added, if necessary.
Particularly, in cultivating the Penicillium sp. (NRRL 18887) under aerobic condition, ordinary aerobic cultivation methods such as, for example, solid culture, culture under aeration and agitation, shaken culture etc. may be favorably utilized. Trace metals, for example, zinc, magnesium, manganese, cobalt, iron and the like should be be added to the fermentation medium since reverse-osmosis grade water must be supplemented.
In carrying out cultivation with aeration and agitation, an anti-foaming agent, e.g., silicon oil, vegetable oils. surfactants, etc. may be suitably employed.
The pH of the medium may be usually within a pH range of 3-9 and preferably within or around neutral range and cultivation temperatures may be usually of 20 0 -30 0 in particular WO 93/10120 PCT/US92/09483 -26about 21°C being preferred.
Cultivation may be continued until Marcfortine A will be substantially accumulated in a culture medium, usually for 20 hours to 240 hours, preferably for 48 hours to 168 hours and, after cultivation, Marcfortine A may be isolated and recovered from a cultured broth by a suitable combination of various method. For example, there may be extraction with an organic solvent, e.g. ether, ethyl acetate or chloroform; dissolution into a more polar solvent, e.g.
acetone or alcohol; removal of impurities with a less polar solvent, e.g. petroleum ether or hexane, adsorptive chromatography with active carbon or silica gel; gel filtration through a column of "sephadex" (available from Pharmacia Co., Ltd, and so on.
Marcfortine B can be prepared from Marcfortine A by biotransformation using a microorgainism in an aqueous nutrient medium containing an assimilable source of carbon and an assimilable source of nitrogen under aerobic conditions. The present invention also provides: Biologically pure cultures of the (Organisms obtained from culture collection as obviously discovered entities) genus Cunninghamella, characterized as species selected from the group consisting of NRRL 1368, 1393 or 3655 and ATCC 8688a.
A method of using members of genus Cunninghamella, characterized as species selected from the group consisting of NRRL 1368, 1393 or 3655, and ATCC 8688a to produce Marcfortine B which comprises: adding Marcfortine A to metabolizing cultures of the cited microorganisms.
By "biotransformation" is meant the use of microorganisms and/or isolated, partially purified enzymes for the converstion of a given substrate into a useful product. H.G. Davies, et al., "Biotransformations in Preparat 'e Organic Chemistry", Academic Press, 1989, p. IX.
By "metabolizing" is meant carrying out the processes of metabolism. A definition of metabolism may be found in A.L. Lehninger, "Principles of Biochemistry", Worth Publ., New York, 1982, p. 333.
By using the new method of the present invention, Marcfortine B is produced by adding Marcfortine A to growing cultures of cited microorganisms. The genus Cunninghamella is preferred for effecting this transformation, including Cunninghamella echinulata subsp. elegans NRRL 1368, Cunninghamella blakesleeana ATCC 8688a Cunninghamella echinulata subsp. elegans NRRL 1393, Cunninghamella echinulata NRRL 3655; the strain NRRL 3655 is particularly preferred.
A subculture of Cunninghamella echinulata subsp. elegans was deposited under the provisions of the Budapest Treaty in the permanent collection of the Northern Region Research Center, ARS; U.S. Dept. of Agriculture; Peoria, Illinois, USA. Its accession number is NRRL 1368. A subculture of Cunninghamella echinulata subsp. elegans was also deposited under the provisions of the Budapest Treaty in the permanent collection of the American Type Culture WO 93/10120 PCT/US92/09483 -27- Collection, Rockville, MD, USA. Its accession number is ATCC 8688b.
A subculture of Cunninghamella blakesleeana was deposited under the provisions of the Budapest Treaty in the permanent collection of the American Type Culture Collection, Rockville, MD, USA. Its accession number is ATCC 8688a.
A subculture of Cunninghamella echinulata subsp. elegans was deposited under the provisions of the Budapest Treaty in the permanent collection of the Northern Region Research Center, ARS; U.S. Dept. of Agriculture; Peoria, Illinois, USA. Its accession number is NRRL 1393. A subculture of Cunninghamella echinulata subsp. elegans was also deposited under the provisions of the Budapest Treaty in the permanent collection of the American Type Culture Collection, Rockville, MD, USA. Its accession number is ATCC 10028b.
A subculture of Cunninghamella echinulata was deposited under the provisions of the Budapest Treaty in the permanent collection of the Northern Region Research Center, ARS; U.S.
Dept. of Agriculture; Peoria, Illinois, USA. Its accession number is NRRL 3655, The use of these microorganisms in the process of the present invention results in the production of Marcfortine B from Marcfortine A.
Marcfortine B is produced when Cunninghamella echinulata subsp. elegans NRRL 1368, Cunninghamella blakesleeana ATCC 8688a, Cunninghamella echinulata subsp.
elegans NRRL 1393 or Cunninghamella echinulata NRRL 3655 is fermented in an aqueous nutrient medium under submerged aerobic conditions in the presence of Marcfortine A.
Typically the microorganism is fermented in a nutrient medium containing a carbon source and a proteinaceous material. Preferred carbon sources include glucose, brown sugar, sucrose, glycerol, starch, comstarch, lactose, dextrin, molasses, and the like. Preferred nitrogen sources include cottonseed flour, corn steep liquor, yeast, autolyzed brewer's yeast with milk solids, soybean meal, cottonseed meal, cornmeal, milk solids, pancreatic digest of casein, distilL.s' solids, animal peptone liquors, meat and bone scraps, and the like. Combinations of these carbon and nitrogen sources can be used advantageously. Trace metals, for example, zinc, magnesium, manganese, cobalt, iron and the like need not be added to the fermentation medium since tap water and unpurified ingredients are used as medium components.
Production of Marcfortine B can be induced at any temperature conducive to satisfactory growth of the microorganism between about 230 and 320 C and preferably at about 28 0
C.
Ordinarily, optimum production of Marcfortine B is obtained in about 1 to 4 days after addition of Marcfortine A to the growing culture, and preferably in about 2 days. The fermentation broth normally remains weakly acid to basic (pH 5.8 9.0) during the fermentation. The final pH is dependent, in part, on the buffers present, if any, and in part, on the initial pH of the culture medium. It is advantageously adjusted to about pH 6.5-7.5, and preferably 7.2, prior to sterilization.
WO 93/10120 PCT/US92/09483 -28- When growth is carried out in shake flasks or large vessels and tanks, it is preferable to use the vegetative form, rather than the spore form, of the microorganism for inoculation to avoid a pronounced lag in the production of Marcfortine B and the attendant inefficient utilization of the equipment. Accordingly, it is desirable to produce a vegetative inoculum in an aqueous nutrient medium by inoculating this medium with an aliquot from a soil or a slant culture. When a young, active vegetative inoculum has thus been secured, it is transferred aseptically to other shake flasks or tanks. The medium in which the vegetative inoculum is produced can be the same as, or different from, that utilized for the production of Marcfortine B, as long as it is such that adequate growth of the microorganism is obtained.
A variety of procedures can be employed to isolate and purify the Marcfortine A from the fermentation broth, for example, by chromatographic adsorption procedures followed by elution with a suitable solvent, column chromatography, partition chromatography, and crystallization from solvents and combinations thereof.
In the preferred recovery process, the Marcfortine B is extracted from the whole beer.
Column chromatography techniques, preferably over silica gel, are used to perform the initial purification. Final purification of marcfortine B is achieved by chrucatography and crystallization from organic solvents.
Procedure A further illustrates the biotransformation process of the subject invention.
The instant compounds of this invention are unexpectedly potent antiparasitic agents against endo and ecto parasites, particularly helminths and arthropods, which cause numerous parasitic diseases in humans, animals, and plants.
Parasitic diseases may be caused by either endoparasites or ectoparasites. Endoparasites are those parasites which live inside the body of the host, either within an organ (such as the stomach, lungs, heart, intestines, etc.) or simply under the skin. Ectoparasites are those parasites which live on the outer surface of the host but still draw nutrients from the host.
The endoparasitic diseases generally referred to as helminthiasis are due to infection of the host with parasitic worms known as helminths. Helminthiasis is a prevalent and serious worldwide economic problem due to infection of domesticated animals such as swine, sheep, horses, cattle, goats, dogs, cats, and poultry. Many of these infections are caused by the group of worms described as nematodes which cause diseases in various species of animals throughout the world. These diseases are frequently serious and can result in the death of the infected animal. The most common genera of nematodes infecting the animals referred to above are Haemonchus, Trichostrongylus, Ostertagia, Nematodirus, Cooperia, Ascaris, Bunostomum, Oesophagostomum, Chabertia, Trichuris, Strongylus, Trichonema, Dictyocaulus, Capillaria, Heterakis, Toxocara, Ascaridia, Oxyuris, Ancylostoma, Uncinaria, Toxascaris, and Parascaris.
Many parasites are species specific (infect only one host) and most also have a preferred site of WO 93/10120 PCT/US92/09483 -29infection within the animal. Thus Haemonchus and Ostertagia primarily infect the stomach while Nematodirus and Cooperia mostly attack the intestines. Other parasites prefer to reside in the heart, eyes, lungs, blood vessels, and the like while still others are subcutaneous parasites.
Helminthiasis can lead to weakness, weight loss, anemia, intestinal damage, malnutrition, and damage to other organs. If left untreated these diseases can result in the death of the animal.
Infections by ectoparasitic arthropods such as ticks, mites, lice, stable flies, homflies, blowflies, fleas, and the like are also a serious problem. Infection by these parasites results in loss of blood, skin lesions, and can interfere with normal eating habits thus causing weight loss.
These infections can also result in transmission of serious diseases such as encephalitis, anaplasmosis, swine pox, and the like which can be fatal.
Animals may be infected by several species of parasite at the same time since infection by one parasite may weaken the animal and make it more susceptible to infection by a second species of parasite. Thus a compound with a broad spectrum of activity is particularly advantageous in the treatment of these diseases. The compounds of this invention have unexpectedly high activity against these parasites, and in addition, are also active against Dirofilaria in dogs, Nematospiroides and Syphacia in rodents, biting insects and migrating dipterous larvae such as Hypoderma sp. in cattle, and Gastrophilus in horses.
The instant compounds are also useful against endo and ecto parasites which cause parasitic diseases in humans. Examples of such endoparasites which infect man include gastro intestinal parasites of the genera Ancylostoma, Necator, Ascaris, Strongyloides, Trichinella, Capillaria, Trichuris, Enterobius, and the like. Other endoparasites which infect man are found in the blood or in other organs. Examples of such parasites are the filarial worms Wucheria, Brugia, Onchocerca, and the like as well as extra intstinal stages of the intestinal worms Strongylides and Trichinella. Ectoparasites which parasitize man include arthropods such as ticks, fleas, mites, lice, and the like and, as with domestic animals, infections by these parasites can result in transmission of serious and even fatal diseases. The instant compounds are active against these endo and ecto parasites and in addition are also active against biting insects and other dipterous pests which annoy humans. The instant compounds when administered orally or parenterally are administered at a dosage rate of from 0.05 to 20 mg/kg of animal body weight.
The instant compounds are also useful against common household pests such as Blatella sp. (cockroach), Tineola sp. (clothes moth), Attagenus sp. (carpet beetle), Musca domestica (housefly) and against Solenopsis Invicta (imported fire ant).
The compounds are furthermore useful against agricultural pests such as aphids (Acyrthiosiphon locusts, and boll weevils as well as against insect pests which attack stored grains such as Tribolium sp. and against inluiature stages of insects living on plant tissue. The WO 93/10120 PCT/US92/09483 compounds are also useful as a nematocide for the control of soil nematodes which may be agr ulturally important.
For use as an antiparasitic agent in animals the instant compounds may be administered intemally either orally or by njection, or topically as a liquid drench or as a shampoo.
For oral administration, the compounds may be administered in capsule, tablet, or drench bolus form or alternatively they can be mixed in the animals feed. The capsules, tablets, and drenches boluses are comprised of the active ingredient in combination with an appropriate carrier vehicle such as starch, talc, magnesium stearate, or di-calcium phosphate. These unit dosage forms are prepared by intimately mixing the active ingredient with suitable finelypowdered inert ingredients including diluents, fillers, disintegrating agents, suspending agents, and/or binders such that a uniform mixture solution or suspension is obtained. An inert ingredient is one that will not react with the instant compounds and which is non toxic to the animal being treated. Suitable inert ingredients include starch, lactose, talc, magnesium stearate.
vegetable gums and oils, and the like. These formulations may contain a widely variable amount of the active and inactive ingredients depending on numerous factors such as the size and type of the animal species to be treated and the type and severity of the infection. The active ingredient may also be administered as an additive to the feed by simply mixing the compound with the feedstuff or by applying the compound to the surface of the feed.
Alternatively the active ingredient may be mixed with an inert carrier and the resulting composition may then either be mixed with the feed or fed directly to the animal. Suitable inert carriers include corn meal, citrus meal, fermentation residues, soya grits, dried grains and the like. The active ingredients are intimately mixed with these inert carriers by grinding, stirring, milling, or tumbling such that the final composition contains from 0.001 to 5.0% by weight of the active ingredient.
The compounds may alternatively be administered parenterally via injection of a formulation consisting of the active ingredient dissolved in an inert liquid carrier. Injection may be either intramuscular, intraruminal, intratracheal, or subcutaneous. The injectable formulation consists of the active ingredient mixed with an appropriate inert liquid carrier. Acceptable liquid carriers include the vegetable oils such as peanut oil, cotton seed oil, sesame oil and the like as well as organic solvents such as solketal, glycerol formal and the like. As an alternative, aqueous parenteral formulations may also be used. The vegetable oils are the preferred liquid carriers. The formulations are prepared by dissolving or suspending the active ingredient in the liquid carrier such that the final formulation contains from 0.005 to 20% by weight of the active ingredient.
Topical application of the instant compounds is possible through the use of a liquid drench or a shampoo containing the instant compounds as an aqueous solution or suspension.
WOi 93/10120 PCT/US92/09483 -31- These formulations generally contain a susoending agent such as bentonite and normally will also contain an antifoaming agent. Formulations containing from 0.005 to 20% by weight of the active ingredient are acceptable. Preferred formulations are those containing from 0.5 to 5% by weight of the instant compounds.
The instant compounds are primarily useful as antiparasitic agents for the treatment and/or prevention of helminthiasis in domestic animals such as cattle, sheep, horses, dogs, cats, goats, swine, and poultry. They are also useful in the prevention and treatment of parasitic infections of these animals by ectoparasites such as ticks, mites, lice, fleas and the like. They are also effective in the treatment of parasitic infections of humans. In treating such infections the compounds of this invention may be used individually or in combination with each other or with other unrelated antiparasitic agents. The dosage of the instant compounds required for best results depelds on several factors such as the species and size of the animal, the type and severity of the infection, the method of administration and the compound used. Oral administration of the instant compounds at a dose level of from 0.005 to 50 mg per kg of animal body weight either in a single dose or in several doses spaced a few days apart, generally gives good results. A single dose of one of the instant compounds normally gives excellent control however repeat doses may be given to combat re-infection or for parasite species which are unusually persistent. The techniques for administering these compounds to animals are known to those skilled in the veterinary field.
The compounds of this invention may also be used to combat agricultural pests which attack crops either in the field or in storage. The compounds are applied for such uses as sprays, dust&, emulsions and the like either to the growing plants or the harvested crops. The techniques for applying these compounds in this manner are known to those skilled in the agricultural arts.
The following examples are provided in order that this invention might be more fully understood; they are not to be construed as limitative of the invention.
Example 1 Production and Isolation of Marcfortine A Seed Fermentation Process: Seed fermentations are inoculated using agar plugs of isolate Penicillium sp. UC 7780 (NRRL 18887) stored over liquid nitrogen. Three plugs are thawed and used as inoculum.
GS-7 is composed of glucose and cottonseed flour (sold under the trademark "Pharmamedia" by Traders Protein, Procter Gamble Oilseed Products Co., Memphis, TN, Unsupplemented tap water is used to hydrate the medium components and the medium is adjusted to pH 7.2 with NH 4 OH. The medium is dispensed into unbaffled closed-system flasks at 300 ml pper 1000 ml flask, and sterilized by autoclaving at 121 0 C for 30 minutes. Each closed-system flask containing 300 ml of GS-7 medium is inoculated with three agar plugs of WO 93/10120 PCT/US92/09483 -32- Penicillium sp. UC 7780 (NRRL 18887) and shaken on a rotary shaker at 250 rpm for 36 hr at 22 0
C.
Secondary Seed Fermentation Process: The mature seed cultures are used as inoculum for the secondary medium at a 0.3% seed rate. The secondary medium is composed of glucose monohydrate (sold as under the trademark Cerelose by C.P.C. International) 25 g, cottonseed flour (sold under the trademark "Pharmamedia") 25 g, MgCl2.6H 2 O 329.8 mg, MnSO 4
.H
2 O 11.4 mg, FeSO 4 .7H,O 3.2 mg, Na 2 MoO 4 *2H20 1.8 mg, CaCI 2 .2HO 367.6 mg, NaCI 84.2 mg, KCI 5.8 mg, ZnSO 4 -7HO 0.1 mg, CoC12-6H 2 0 0.1 mg, CuSO 4 *5H 2 0 3.1 mg, and silicone antifoam (sold under the trademark SAG-471 Antifoam) 0.5 ml per liter of reverse-osmosis grade water. Medium components sufficient for 200 liters of secondary seed medium are hydrated in reverse-osmosis grade water to a q.s. volume of 190 liters in a 250-L fermentor. After formulation, the pH of the medium is adjusted to pH 7.2 with NH 4 OH, and then the medium is sterilized at 121 0 C for 30 minutes.
Two closed-system flasks of the mature primary-seed culture are used as inoculum at a 0.3% seed rate. The secondary seed culture is incubated at at 22 0 C, with 125 slm aeration, 5 psig backpressure, and 250 rpm for 36 hours.
Production Fermentation Process: The production medium is composed of beet molasses 50 g, fish meal (sold under the trademark Menhaden Select Fish Meal) 16 g, yeast extract (sold under the trademark Fidco) g, MgCI 2 .6H 2 0 329.8 mg, MnSO 4
*H
2 0 11.4 mg, FeSO 4 *7H 2 O 3.29 mg, Na 2 MoO 4 .2H 2 0 1.8 mg, CaC 2 1,2H 2 0 367.6 mg, NaCI 84.2 mg, KCI 5.8 mg, ZnSO 4 -7H0O 0,1 mg, CoC12*6H 2 0 0.1 mg, CuSO 4 *5H0O 3.1 mg, and silicone antifoam (sold under the trademark SAG-471 Antifoam) ml per liter of reverse-osmosis grade water.
Medium components sufficient for 5,000 liters of medium are hydrated in reverseosmosis grade water to a q.s. volume of 4,700 liters in a 5,000 L fermentor. After formulation, the pH of the medium is adjusted to pH 7.0 with KOH, and then the medium is sterilized at 123 0 C for 30 minutes. The mature secondary-seed culture is used as inoculum at a 1.0% seed rate. The culture is incubated at 22°C, with 2,500 slm aeration, 5 psig backpressure, and 250 rpm for 96 hours.
Isolation of Marcfortine A: The 4900 L fermentation volume is harvested by passing through a high shear mixer to the harvest vessel. Following transfer, 4% wt./v. of diatomaceous earth and 1/2 volume of methylene chloride are added. The harvest solution is then filtered using a filter press. The filter cake is washed 2 times with a 10% volume of methylene chloride, The filtrate obtained is decanted to remove the water (aqueous) phase. The remaining product-rich methylene chloride phase is then concentrated to a volume of 44 L. The 93/10120 PCT/US92/09483 -33concentrate is then polished using a 20% concentrate volume 9 L)of methylene chloride and diatomaceous earth over a filter.
The 53 L polished concentrate is further purified to separate Marcfortine A from other components by silica gel chromatography and crystallization.
Before chromatography, the polished concentrate is divided into four approximately equal aliquots. Each aliquot is chromatogriphed over a newly packed 9" diameter column prepared from 25 Kg of dry silica gel (bed volume 59 The loaded columns are eluted with 120 L of 10% acetone in methylene chloride, 120 L of 20% acetone in methylene chloride, 120 L of 30% acetone in methylene chloride, 160 L of 40% acetone in methylene chloride, and 130 L of acetone collecting the 30 and 40% eluates 20 L fractions. Eluates are monitored by TLC, usm. for example a solvent system comprised of 6% isopropanol and 0.3% ammoniumn hydroxide in methylene chloride to develop Whatman LK6DF silica gel plates. Fractions of Marcfort;' (contai. .ag a small amount of Mar:fortine D which co-chromatographs with D) are crys; J from acetone. Appropriate fractions 40-100 L) are concentrated under reduced pressure to a volume of approximately 5 L. The solution or light slurry) is then transferred to a rotatory evaporater and conce l:on continued under reduced pressure. Several L portions of acetone are added during t. surse of the concentration until the methylene chloride is completely displaced. The resulting acetone slurry (approximately I L volume) is refrigerated overnight, and the crystals of Marcfortine A are collected and washed with several small portions of cold acetone, and dried under vacuum. Such crystals may be contaminated with several percent of Marcfortine D. Repeated recrystallization from methylene chloride/acetone (displacing methylene chloride as described) affords pure Marcfortine A.
Example 1A Production and Isolation of Marcfortines A and C.
Primary Seed Fermentation Process: Seed fermentations are inoculated using agar plugs of isolate Penicillium sp. UC 7780 (NRRL 18887) stored over liquid nitrogen. Three plugs are thawed and used as inoculum for 100 ml of GS-7 seed medium. GS-7 is composed of glucose and cottonseed flour (sold under the trademark "Pharmamedia" by Traders Protein, Procter Gamble Oilseed Products Co., Memphis, TN, each added at a concentration of 25 g/L of tap water. After formulation, the pH of GS-7 is adjusted to 7.2 using NH 4 0H. The medium is autoclaved in 100 ml volumes in 500 ml unbaffled fermentation flasks for 30 min. Sterile GS-7 is inoculated as described above and shaken at 250 rpm for 35-58 hr at 23 0
C.
Production Fermentation Process (shaker flask): The mature seed cultures are used as inoculum for the production medium at a 1% seed rate. The production medium is composed of glucose 45 g, enzymatically digested casein (sold under the trademark Peptonized Milk Nutrient by Sheffield Products, Norwich, WO 93/10120 PCT/US92/09483 -34g, yeast extract (sold under the trademark BACTO Yeast Extract Code: 0127 by Difco Laboratories, Detroit, MI) 2.5 g per liter of tap water. After formulation, the pH of the production medium is adjusted to 7.0 using potassium hydroxide. This medium is then autoclaved for 30 min in 100 ml volumes- contained in 500 ml baffled fermentation flasks.
Sterile production medium is inoculated as described above, and shaken for 7-14 days at 250 rpm at 21 0
C.
Production Fermentation Process (Labraferm tanks): The mature seed cultures are used as inoculum for the sterile production medium at a seed rate. The production medium is described above. After pH adjustment to 7.0 using KOH, 10 L of this medium are autoclaved for 90 min in 12 L Labraferm tanks (New Brunswick Scientific Co., Inc.). The tanks are inoculated at a 0.5% seed rate and stirred at 500 rpm at for 5-9 days. The air flow rate is maintained between 10-15 L/min.
Isolation of Marcfortines A and C: Whole fermentation broth (35 1) is macerated at low speed in a large commercial Waring Blender and then blended with an equal volume of methylene chloride. The mixture is stored overnight under refrigeration and then subjected to centrifugation to break the emulsion. The resulting clear methylene chloride layer is drawn off and evaporated under reduced pressure. A concentrated solution of the residue (37.4 g) in methylene chloride is applied to a column of silica gel (1Kg) slurry packed in methylene chloride. The column is eluted with increasing concentrations of acetone in methylene chloride (10 20%, 30%, 40%, and 50% acetone).
Fractions are monitored by TLC and appropriate fractions evaporated and crystallized from acetone to give Marcfortine A and Marcfortine C.
Example 1B Production and Isolation of Marcfortines A and C.
Seed Fermentation Process: Seed fermentations are inoculated using agar plugs of isolate Penicillium sp. UC 7780 (NRRL 18887) stored over liquid nitrogen. Three plugs are thawed and used as inoculum for 100 ml of GS-7 seed medium. GS-7 is composed of glucose and cottonseed flour (sold under the trademark "Pharmamedia" by Traders Protein, Procter Gamble Oilseed Products Co., Memphis, TN, each added at a concentration of 25 g/L of tap water. After formulation, the pH of GS-7 is adjusted to 7.2 using NH 4 OH. The medium is autoclaved in 100 ml volumes in 500 ml unbaffled fermentation flasks for 30 min. Sterile GS-7 is inoculated as described above and shaken at 250 rpm for 35-58 hr at 23 0
C.
Production Fermentation Process (Shake Flask): The mature seed cultures are used as inoculum for the production medium at a 1% seed rate. The production medium is composed of glucose 20 g, glycerol 15 ml, cottonseed flour (sold under the trademark "Pharmamedia" by Traders Protein, Procter Gamble Oilseed WO 93/10120 PCT/US92/09483 Products Co., Memphis, TN, 20 g, soybean meal 10 g, and K2HPO 4 3 g per liter of tap water. After formulation, the pH of the production medium is adjusted to 6.8 using potassium hydroxide. This medium is then autoclaved for 30 min in 100 ml volumes contained in 500 ml baffled fermentation flasks. Sterile production medium is inoculated as described above, and shaken for 7-14 days at 250 rpm at 21°C.
Production Fermentation Process (Labraferm tanks): The mature seed cultures are used as inoculum for the sterile production medium at a seed rate. The production medium is described above. After pH adjustment to 7.0 using KOH, 10 L of this medium are autoclaved for 90 min in 12 L Labraferm tanks (New Brunswick Scientific Co., Inc.). The tanks are inoculated at a 0.5% seed rate and stirred at 500 rpm at 0 C for 5-9 days. The air flow rate is maintained between 10-15 L/min.
Isolation of Marcfortines A and C: Whole fermentation broth (35 1) is macerated at low speed in a large commercial Waring Blender and then blended with an equal volume of methylene chloride. The mixture is stored overnight under refrigeration and tl subjected to centrifugation to break the emulsion. The resulting clear methylene chloride 1 is drawn off and evaporated under reduced pressure. A concentrated solution of the residue g) in methylene chloride is applied to a column of silica gel (1Kg) slurry packed in methylene chloride. The column is eluted with increasing concentrations of acetone in methylene chloride (10 20%, 30%, 40%, and 50% acetone).
Fractions are monitored by TLC and appropriate fractions evaporated and crystallized from acetone to give Marcfortine A and Marcfortine C.
Example 2 18-thiomarcfortine A (Compound #2) A solution of 30 mg marcfb.tine A and 22 mg of 2,4-bis(methoxyphenyl)-1,3-dithia-2,4diphosphetane-2,4-disulfide (Lawesson's reagent) in 5 ml of toluene is refluxed under nitrogen for 18 hours. The mixture is coo ed and the solvent removed under reduced pressure. The residue is subjected to preparative thin layer chromatography on silica gel plates using acetone in methylene chloride as the eluent to give 18-thiomarcfortine A, mp 258-260 °C.
FAB-MS 494(M +H) Following the general procedure of Example 2 but employing the appropriate marcfortine starting material, there are prepared the following other 18-thiomarcfortine compounds: l-methyl-18-thiomarcfortine A 1-benzyl-18-thiomarcfortine A 1-ethyl-18-thiomarcfortine B 1-benzyl-18-thiomarcfortine B 18a-ethyl-18-thiomarcfortine B WO 93/10120 WO 9310120PCT/US92/09483 -36- 1 8a-benzyl- 18-thiomarcfortine B I 8a-methoxyethoxymethyl-1I8-thiomarcfortine B 18a-allyl-18-thiomarcfortine B I 8a-propargyl-l18-thiomarcfortne B 1, 18a-bis-ethyl- 1 8-thioinarefortine B 1, 1 8a-bis-benzyl- 1 8-thiomarcfortine B 1 8a-ethyl-24-methoxy- 18-thiomarcfortine B I 8-thiomarcfortine B (Compound #2A) FAB-MS 480(M'+H) 1 8a-ethyl-24,25 dihiydro- 18-thiomarcfortine B l-propyl-1 8-thiomarcfortine C I -benzyl-1I8-thiomarcfortine C 1 Sa-propyl- 18-thiomarcfortine C 1 8a-benzyl- 18-thiomarcfortine C 1 8a-methoxyethoxymethyl-18-thiomarcfortine C 18a-aIlyl-18-thiiomarcfortine C 1 Sa-propargyl- 18-t~homarcfortine C 1,1 8a-bis-propyl- 18-thiomarcfortine C 1,1 8a-bis-tienzyl- 18-thiomarcfortine C 1 8-thiomarcfortine C 1-palmitoyl-1 8-thiomarcfortine- A 1 -decanoyl- 18-thiorarcfortine A 1 -decanoyl-1I8-thiomarcfortine B Example-3 24,25 dihydro-18-thiomarcfortine A A mixture consisting of 15 mg of 5% palladium on carbon and 1 8-thiomarcfortine A mg, 0.06 mmol) in 1 ml of methanol is stirred vigorously under an atmosphere of hydrogen for 45 minutes. The reaction mixture is filtered through Ceite® and the filtrate is evaporateci under vacuum. Preparative layer chromatography of the residue on a 0.5 mm silica gel plate eluted with 3% methanol in methylene chloride affords 24,25-dihydro iS-thiomarefortine A.
Example 4 24,25-dibromo-24,25-dihydro 18-thiomarcfortine A A solution of bromine in chloroform (0.4 ml of 12M solution, 0.048 mmol) is added drepwise to a cold (ice bath) solution of 18-thlomarcfortine A (20 mg, 0.04 mmol) in 2 ml of chloroform. The Yesulting yellow solution is stirred at room temperature for 15 minutes then evaporated unde-r a steamn of nitrogen. Preparative layer chromatography of the residue on a 0.5 mm silica gel plate eluted with 2% methanol in methylene chloride affords 24,25-dibromo- 24,25-dihydro- 18-thiomarcfortine A.
WO 93/10120 PCT/US92/09483 -37- Example 5 24-methoxy 24,25-dihydro 18-thiomarcfortine B A solution of bromine in chloroform (0.6 ml of 0.12M solution, 0.072 mmol) is added dropwise to a cold (ice bath) solution of 18-thiomarcfortine B (28 mg, 0.06 mmol) in 2 ml of chloroform. The resulting yellow solution is stirred at room temperature for 10 minutes then at 0 C for 20 minutes then evaporated under a stream of nitrogen. The yellow solid residue is dissolved in 2 ml of methanol then 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 0.015 ml, 0. .0 mmol) is added. The solution is stirred at room temperature for 90 minutes then evaporated under vacuum. Preparative layer chromatography of the residue on a 1.0 mm silica gel plate eluted with 2% methanol in methylene chloride affords 24-methoxy-25-bromo-24,25-dihydro-18thiomarcfortine B. The 24-methoxy 25-bromo-24,25-dihydro-18-thiomarcfortine B thus obtained is dissolved in 2 ml of dry toluene then tributyltin hydride (0.12 ml, 0.45 mmol) is added. The solution is stirred at 1000 C. for 16 hours then evaporated under vacuum. Preparative layer chromatography of the residue on a 2.0 mm silica gel plate eluted~ with 2% methanol in methylene chloride affords 24-methoxy 24,25-dihydro-18-thiomanrfortine B.
Example 6 24-propoxy 24,25-diIydro-18-thiomarcfortine A Substitution of propanol for methanol in the procedure described above for 24-methoxy 24,25-dihydro-18-thiomarcfortine B (Example 5) and application of the modified procedure to mg of 18-thiomarcfortine A affords an oily residue. Preparative layer chromatography of the crude produce on a 0.5 mm silica gel plate eluted with 2% methanol in methylene chloride affords 24-propoxy-24,25-dihydro-18-thiomarcfortine A.
Example 7 l-(dimethylcarbamoyl)-18-thiomarcfortine A Potassium hydride (50 mg of a 25% oil dispersion) is added to a solution of 18-thiomarcfortine A (15 mg, 0.030 mmol) in 1 ml of dry tetrahydrofuran. The solution is stirred at room temperature for 2 hours then dimethylcarbamoyl chloride (0.028 ml, 0.30 mmol) is added. The mixture is stirred at room temperature for 18 hours then partitioned between aqueous sodium bicarbonate (1 ml) and methylene chloride (1 ml). The layers are separated and the aqueous layer extracted with methy!lne chloride (2 ml). The combined extracts are dried with magnesium sulfate, filtered, and evaporated under vacuum. Preparative layer chromatography of the residue on a silica gel plate eluted with 25% acetone in hexane afOards 1-(dimethylcarbamoyl)-18-thiomarcfortine A.
Following the general procedure of Example 7 but employing the appropriate marcfortine starting material, there are prepared the following other 18-thiomarcfortine compounds: 1-acetoxymethyl-18-thiomarcfortine A 1-diethoxyphosphoryl-18-thiomarcfortine A 1-cyclopropylcarbonyl- 18-thiomarcfortine A WO 93/10120 WO 3/1120PCT/US92/09483 -38- I -dimethylsulfamoyi-18-thiomarcfortine A l-(1-l ipetidiny)thiocar'onyI-1 8-thiomarcfortine, A I -saccinoyl- 18-thlomarcfortine A I -(2,4-dinitrobenzenesulfenyl)- 18-thiomarefortinie A '-(4-morphiolinosulfenyl)-1I8-thiomarcfortine A I -(p-toluenesulfonyl)-1I8-thiomarcfortine A I -acetyl-1I8-thiomarcfortine A I -methoxycarbonyl-1I8-thiomarcforth, e A I -(p-toluenesulfonyl)- 1 Sthiomarcfortine B 1 -(2,4-dinitrobenzeiie sulfonyl)-1I8a-ethyl-24-methoxy-24,25-dihydro I -(p-bromobenzene sulfonyl)- 18-thiomarcfortine C I -propionyl-1I8-thiomarcfortine C I -(4-carbethoxy- 1,3-thiazolidinn3-yl)carbonyl-1 8-thiomarcforrine A .FxaMple 8 1-benzyl-18-thio-marcforrine A Potassium hydride (75 mg of a 25% oil dispersion) is added to a solution of 18-thiomarcfortine A (15 mg, 0.030 romol) in I ml of dry tetrahydrofuran. The solution is stirred at room temperature for 2 hours then benzyl b.romide (0.025 ml, 0.21 mmol) is added. The mixture is stirred at room temperature for 3 hours then partition.-d between water (I ml) and methy).ne chloride (I ml). Trie layers are separated and the~ aqueous layer extracted with methylene chloride (2 ml). The combined extracts are dried with magnesium sulfate, filtered, anid evaporated under vacuum. Preparative layer chromatography of the residue on a 1.0 mm silica gel plate eluted with 3% methanol in methylene chloride affords I N-bto,1-l8-thiomarcfortine A.
Following ihe; general procedure of Example 8 but employing the appropriate marcfortine starting material, there are prepared the following other I .8-thiomarcfortine compounds: 1,1 8a-bis-ethyl- 18-thiomarcfortine B; I -allyl- I -thiomarcfOrtine A; 1-methoxymethyl-l 8-thiomrarcfortine A; I-propargyl-1 8-thiomarcfortine A; I -benzyl- 18a-ethyl-l 8-thiomarcfoniine B; Example 9 Haemnonchus confortusITrichostrongylu 91albriformislJird Assay: This in vivo assay utilizes jirds infected with two impon. .t target parasites of rumninants, H. contortus and T. colubriformis (anthelmintic-sensitive or -resistant worvs can be used).
Initially, activity is asaessed only against H. contortus as described in G.A. Conder et al., 3.
Parasitol. 76, 168-170 (1990), while follow-up studies examine activity against both species of WO 93/10120 WO 3/1120PCTr/US92/09483 -39parasites using the technidques outlined in G.A. Conder et al., J. Parasitol. 77, 62 1-623 (1991), The activity is set forth in Table II.
Examp~le Following the general procedure of Example 2 bmt employing the appropriate paraherquamide/thoparaherquamide starting material, there are prepared the following other 1 8-thioparaherquamnide compounds: 1 8-thioparaherquamnide I -methyl-I 8-thioparaherquamide I -ethyl-I 8-thioparaherquamnide 1 -benzyl- 1 8-thioparaherquamide I -palmitoyl- 1 8-thioparaherquamide I -decwi oyl-1I 8-thioparaherquamide Examnle Following the general procedure of Example 7 but employing the appropriate thioparaherquamnide starting material, there aire prepared the following other 1 8-thioparaherquariide compounds: 1 -acetoxymethyl- 18-thiomarcflortine 1 -diethoxyphosphoryl- I 8-thioparaherquainide I -cyclopropylcarbonyl-1 8-thioparaherquamide 1 -dimethylsulfamoyl-I 8-thioparaherquamide I -piperidinyl)thiocarbonyl- 1 8-thioparaherquamide I -succinoyl-1 8-thioparaherquamide 1 -(2,4-dir~obenzenesulfenyl).' 1 8-thioparaherquamide 1-(4-morphnoiinosulfenyl)- I 8-thioparaherquamide 1 -(p-toluenesulfonyl)- 1 8-thioparaherquamnide I -acetyl- 1 8-thioparaherquamide I -methoxycarbonyl- 1 8-thioparaherquamide Example 11I Following the general procedure of Example 8 but employing the appropriate paraherquamide starting material, there are prepared the following other 18-t~hoparaberquamnide compounds: 1, 18a-bis-ethyl- 1 8-thioparaherquamide; I -ailyl-1 8-thioparaherquamide; I -methoxymethyl-1 8-tlioparahemquamide; 1 -propargyl-l 8-thioparaherquamide; 1 -benzyl- 1 ga-ethyl- I 8-thioparaherquamide.
WO 93/10120 WO 93/0120 CI'/US92/09483 Example 12 1-pyrrolidir carbonyi-marcfortine A, Cpd #12 Potassium hydride (50 mg of a 25% oil dispersion) is added to a solution of marcfortine A (15 mg, 0.030 mmol) in 1 ml of dry tetrahydrofuran. The solution is stirred at room temperature for 2 hours then pyrrolidinylcarbonyl chloride (40 mg, 0.30 minol) is added.
The mixture is stirred at room temperature for 18 hours then partitioned between 5% aqueousodium bicarbonate (1 ml) and methylene chloride (I ml). The layers are separated and the aqueous layer extracted with methylene chloride (2 ml). The combined extracts are dried with magnesium sulfate, filtered, and evaporated under vacuum. Preparative layer chromatography of the residue on a silica gdI plate eluted with 25% acetone in hexane affords 1-pyrrolidinyl carbonyl-marcfortine A. 'H NMR(CDC 3 5 0.83(s,3H), l.21(s,3H), 1.41 l.45(s,3H), 1.3-2.4(m,17H), 2.38(d, 1H), 2.6-2.8(m, 2H), 2.84(d,1H), 3.00(s,3H), 3.4-3.6(m,4H), 3.72(d, IH), 4.85(d,1H), 6.4 1(d, 1H), 6.84(s,2H) Following the general procedure of Example 12 but employing the appropriate marcfortine starting material and carbonyl chloride, there are prepared the following other marcfortine compounds: 1 -diethoxyphosphoryl-marcfortine A l-cyclopropylcarbonyl-marcfottine A, Cpd #12A. 'H NMR(CDC 3 6 0.82(s,3H), 1 .05(s.3H), I .44(2s,6H), 0.9-1.7(m,8H), I .7-3.0(m,1 1H), 3. 14(s,3H). 3.80(d, 1H),, 4.84(d,IH), 6.28(d,IH), 6.88(s,211) 1-(1-piperidinyl)thiocarbonyl-marcfortine A 1-succinoyl-marcfortine A 1-phenoxycarbonyl-marcfoftie A, Cpd #12B. 1 i'I i TW(CDCI 3 6 0,83(s,3H), 3.70(d, 1H), 4.83(d, 11), 6.27(d, IH), 6.90(s,2H), 7.2-7.5(m,5H) 1 -(2,4-dichlorophenoxycarbonyl)-1I8a-allyl-marcfortine B I -(4-oxopentanoyl)- 18a-methyl-marcfortine C 1-palmitoyl-marcfortine A, Cpd #12C FAB-MS 716(M-+H) 1-(4-nitrophenoxycarbonyl)-marcfortine A, Cpd #12D. NMR(CDC 3 6 0,90(s,3H), 1.09(s,3H), 1.3-1.5(m,1H), 1.44(s,3H), 1.47(s,311), 1.5-2.9(m,13H), 3.14(s,3H), 3.70(d,IH), 4.86(d,1H), 6.23(d,1H), 6.93(ABq,2H), 7.48(d, 2H), 8.32(d,2H) I 1-piperidiniecarbonyl)-marcfortine A, Cpd #12F FAB-MS 589 HRMS m/z (M 4 C3,H,,NO 5
+H
1 calc 589.3390, obsd 589.3398 1-[[4-(ethoxycarbonyl)piperazin-1-yIlcarbonyll-marcfortine A, Cpd #12G FAB-MS 662 684 HRMS WO 93/10120 WO 9310120PCr/US92/09483 -41-
C
36
H
47
N
5 0 7 calc 662.3553, obsd 662.3569 1-[[4-(benzy1)piperazin- 1-yljcarbonylj-marcfortine A, Cpd #12H 'H NMIR (300 mHz, CDCI 3 0.80 (sOH), 1.26 1.41 1.45 (s,3H), 1.2-2.9 (in, 2.98 3.4-3.8 6.41 6.84 (s,211), 7.2-7.5 FAB-MS 680 702 (M++Na) 1 ,3-bernzodioxol-5-ylmethyl)piperazin-1 -yl]carbonyl]-marcfortine A, Cpd #121 'H NMR (300 mHz, CDCI 3 0.80 1.00-2.60 (m,ISH), 1.2 1.41 1.45 2.63 (t,IH),2.84 2.98 3.45-3.80 (m,8H), 4.85 5.95 6.41 6.70-6.90 FAB-MS 724 1-f [4-(methyl)piperazin-1-ylllcarbonylll-marcfortine A, Cpd #1 2J 'H NMR (300 mHz, CDCI 3 0.82 1.26 1.40-2.72 (m,20H), 1,41 1.44 2.82 2.99 (s,31-fl 3.40-3.81 4.85 (d,1H), 6.41 6.84 FAB-MS 604 (M 4 HRMS m/z (M+H,
C
34
H
45
N
5 0 5 calc 604.3499, obsd 604.3506 1 ,3-dihiydro-1I,3-dioxo-2H-isoindo]-2-yl)-marefortine A, Cpd #12K FAB-MS 665 (W-i-H) 1 -[[4-(pyridin-2-yl)piperazin-1 -yl]carbonyl]-marcfortine A, Cpd #12L 'H NMIR (300 mHz, CDCI 3 0.84 1.10-2.30 (m,12H), 1.41 1.44 2.40 2.67 2.85 3.00 3.45-3.85 (m,9H), 4.85 6.40 6.60-6.70 6.85 7.52 8.15-8.22 FAB-MS 667 (M+H) 1 [4-(phenyl)piperazin- 1-yl]carbonyl]-marcfortine A, Cpd #12M 'H NMR (300 mHz, CDCI 3 0.84 1.20-2.50 (m,IIH), 1.22 1.42 1.45 2.67 2.85 3.00 3.,10-3.91 4.85 6.40 6.85 6.90-7.00 7.25-7.38 (in,2H) 1 -1j4-(cldorocarbonyl)piperazin-1I-yllcarbonyllj-marcfortine A, Cpd #1 2N 'H NMIR (300 mHz, CDCI 3 0.74 1. 18 1.30-2.65 (m,20H), 2.74 2.81 3.48-3.90 4.30 6.32 6.80 (s,2H).
FAB-MS 652 (M+-iH) 10"-(1I,4-dicarbonylpiperazine)bis(6',7',8',9', 10', 10'a-hexahydro- 12'-pentamnethyl)-[2'S-[2'.alpha.,3'a.alpha.,9'a.alpha.,1I0(21"R*,3 aS*9 aS*, -Spiro[4H,8H-[I1 ,4]dioxepinoll2,3-glindole-8,2'(3'H)-[ 1H,4H-3a,9a](iminomethano)cyclopentalbI quinolizin]-9,1 1'(10H)-dione, Cpd #120 FAB-MS 1093(M++H) WO 93/10120 W093/0120PCT/US92/09483 -42- I -[I[4-(phenyl)piperidin- 1-yI]carbonyl]-marcfortine A, Cpd #1 2P 1-H NMR (300 mHz, CDCI 3 0.76 1.10-2.30 (m,13H), 1.14 1.35 1.38 2.33 2.52-3.00 (m,811), 3.00-3.20 3.50-3.75 4.05 (d,111), 4.33 4.79 (dd,1H), 6.35 (dd,1H), 6.78 (s,2H), 7.10-7.30 (m,511). FAB-MS 665 (M+H) 1-[[4-(dimethyl)piperidin-1-yllcarbonyl]-marcfortine A, Cpd #12Q 'H NMR (300 mnlz, CDCI 3 0.75 0.90-1.95 (m,13H), 0.92 0.96 1.14 1.34 1.37 2.08 2.20 (brs.,1H), 2.33 (d,11H), 2.60 2.77 1H1), 2.92 3.45-3.75 4.77 (d,1H), 6.33 6.77 FAB-MS 617 1 -[[4-(5-chloropyridazin-3-yI)pipzerazin- 1-yIlcarbonyl]-marcfortine A, Cpd #12R 'H NMR (300 mHz, CL)C1 3 0.83 1.20-2.30 I IfH), 1.22 1.42 1.44 2.45 2.65 2.83 2.99 3.60-3.85 4.86 6.40 6.86 692 7.29 (d,1H).
FAB-MS 702 1 -[[4-(4-cliorophenyl)piperazin- 1-ylllcarbonylll-marcfortine A, Cpd #12S 'H NMR (300 mHz, CDC1 3 0.84 1.20-2.30 (m,914), 1.26 1.42 1.45 0s3H), 2.40 2.60-2.75 2.85 3.00 (s,3H), 3.15-3.30 3.49 3.52-3.84 4.85 6.39 (d,IH), 6.80-6.90 7.23 (d,2H).
10,1 1,4-dioxo-2-butene)bis(6',7',8',9', 10', 10'a-hexahydro- 12'-pentametliyl)-[2'S- [2'.alpha.,3'a.alpha.,9'a. alpha., I I 0'abeta.jj-Spiro[4H,8H- 1,4]dioxepino[2,3-gjindole-8,2'(3'H)-[ 1 H,4H-3a,9a] (iminomethano)cyclopenta[b] quinolizin]-9, 1 1'(1OH)-dione, Cpd #12T FAB-MS 1035(M'-iH) 1 -[[4-(tichloroethoxycarbonyl)piperazin- 1-yIlcarbonyl]-marcfortine A, Cpd #12U 'H NNM (300 mHz, CDC 3 0.75 1.1-1.85 1.15 1.34 1.34 1.92 2.00-2.40 2.34 2.55 (t,2H), 2.76 2.91 3.42-3.75 4.65-4.77 4.79 6.33 6.79 FAB-MS 764 HRMS m/z
C
36 H'4CI 3
N
5
O
7 caic 764.2384, obsd 764.2366 I -[[4-(3-trifluoromethyl-tiadiazol- 1-yI)piperazin-1I-yI]carbonyl]-marcfortine A, Cpd #12V NMR (300 mHz, CDC1 3 0.83 1.26 (s,3H1), 1.30-2.30 11H), 1.42 1.45 2.42 2.55-2.72 2.84 2.99 (s,3H), 3.60-3.90 4.87 6.30 6.88 FAB-MS 742 WO 93/10120 WO 9310120PCT/US92/09483 -43- HRMS m/z C 36
H
42
F
3
N
7 calc 742.2998, obsd 742.3038 Example 13 J-acetoxymnethyl-marcfortine A, Cpd #13 Sodium hydride (47 mag of a 60%7 oil dispersion) is added to a solution of marcfortine A (850 mng, 1.78 rnmol) in I ml of dry tetrahydrofuran. The solution is stirred at room temperature for 3 hours then methyl bromoacetate (280 mg, 1.82 mmul) is added. The mixture is stirred at room temperature for 4 hours then additional methyl bromoacetate (280 mg. 1.82 mmol) is added. The mixture is stirred at room temperature for 8 hours then partitioned between sodium bicarbonate (1 ml) and methylene chloride (1 ml). The layers are separated and the aqueous layer is extracted with methylene chloride (2 ml). The combined extracts are dried with magnesium sulfate, filtered, and evaporated under vacuum. Column chromatography of the residue on silica gel plate eluting first with 5% then 10% acetone in methylene chloride affords 1 acetoxymethyl-marcfortine A as a colorless oil: 'H NMIR (CDC 3 5 0.75 (3H, 1.08 (3H, 1.43 (3H, 1.45 (3H, 2.05 3.12 (3H, 4.86 (IH, d, J 5.85 (IH, d, J 5.96 (1IH, d, J 6.26 (1IH, d, J 6.77 (1IH, d, J 6.86 (11H, d, J1 7).
Following the general procedure of Example 13 but employing the appropriate marcfortine starting material, there are prepared the following other i-arcfortine compounds: 1-t-butyiyloxymethyl-raarcfortine A 1 -benzoyloxymethyl-marcfortine A 1 -acetoxymethyl-1I8a-benzy!-marcfortine B 1-3,4-dichlorobenzoyloxy-marcf-ortine C Example 14 1-(2,4-dinitrobenzeneslfenyl)-marcfortine A, Cpd #14 Marcfortine A (75 mg, 0. 15 mmol) is added to a suspension of potassium hydride (180 mg, 1.50 mmol, 35 wt% in mineral oil) in THF (6 mL) at 5*C, followed by 2,4-dinitrobenzene -sulfcnyl chloride (73 mg, 0.30 mmol). The reaction mixture is then stirred for 3h at 5 0 C. After diluting with H 2 0 and extracting with CHCI 3 the crude product is chromatographed. on a 1 mmn silica gel preparative thin layer plate eluting with 30% Acetone in methylene chloride to yield 1-(2,4-dinrrobenzenesulfenyl)-marcfortine A (90 rag, 89.1%) as a yellow solid, rap 155-160'C (dec). 'H NMR (CDCI 3 5 0.86-0.97 (in,3H), 1.13-1.45 1.56-1.78 1.86 2.00 2.15 2.30 2.45 2.67 (bd,IH), 2.79 2.97 3.08 3.74 4.76 (bd,lH), 5.85 6.87 6.97 7.28 7.62 8.36 (dd,1H), 9.16 (d,1H).
Following the general procedure of Example 14 but employing the appropriate marcfortine starting material, there are prepared the following other marcfortine compounds: 1-(4-morpholinosulfenyl)-marcfortine A, Cpd #14A FAB-MS; m/e, 595(m+-iH) 1-(trichloromethylsulfenyl)-marcfortine A, Cpd 01413 WO 93/10120 PCT/US92/09483 -44- FAB-MS; m/e, 626, 628, 630 1-(methoxycarbonylsuffenyl)-marcfortine A, Cpd #14C) FAB-MS; m/e, 568 I-(berizenesulfenyl)-marcfortine A. Cpd #14D FAB-MS; m/e, 586 (M +H) 1-trichloromethylsulfenyl- 18a-ethyl-marcfortine B 1-ethoxycarbonylsulfenyl-18a-methyl-marcfortine C Example 15 1-(4-morpholinecarbonyl)-marcfortine A N-oxide, Cpd 1-(4-Morpholinecarbonyl)-marcfortine A (Cpd #12E, 15 mg) is treated with 3-chloroperoxybenzoic acid (15 mg) in methylene chloride (2 ml) at 0°C for 10 minutes. The mixture is partitioned between 5% sodium bicarbonate (2 ml) and methylene chloride (2 ml).
The layers are separated and the aqueous layer is extracted with methylene chloride (2 ml). The combined extracts are dried with magnesium sulfate, filtered and evaporated under vacuum. The residue is chromatographed on a 0.5 mm silica gel preparative thin layer plate eluting with methanol in methylene chloride to yield the title compound (10 mg) as a solid. FAB-MS; 607(M +H) Example 16 1-(2-tetrahydrofuranyl)-marcfortine A, Cpd #16 Carbonyldiimidazole (50 mg) is added to a solution of marcfortine A (60 mg) in 6 ml of tetrahydrofuran. The mixture is heated to reflux and potassium hydride (120 mg) is added. The resulting mixture is stirred under refluxing for 1 hour. The precipitate is filtered off and the filtrate is concentrated. The residue is chromatographed on a silica plate, eluting with acetone in methylene chloride to yield the title compound (30 mg) as a solid. HRMS (FAB); m/e(M C 3 2
H
41
N
3 0 5 calc. 548.3124, obsd. 548.3086 Example 17 1-(4-morpholinecarbonyl)-marcfortine A, Cpd#12E Marcfortine A (1 g, 2.1 mmol) in THF (25 mL, distilled from sodium metal and benzophenone) and potassium hydride (35 weight 0.5 g, 4.4 mmol) are stirred for 0.5 h at room temperature under nitrogen. Morpholinecarbonyl chloride (1 mL, 8.4 mmol) is then added via syringe. After 3 hr of stirring at room temperature the turbid reaction mixture is cooled in an ice water bath and quenched with dropwise addition of cold saturated potassium carbonate solution (5 mL). The mixture is then diluted with water (75 mL) and extracted into CH 2 CIl (175 mL). The organic layer is dried over magnesium sulfate, filtered and concentrated to give the crude material which contains oil. Silica gel chromatography to 5% MeOH/CHCIy) gives 1-morpholinecarbonyl-marcfortine A. (Silica gel chromatography on the chromatotron MeOH/CHCI 2 is a better method of purification for 1-morpholinecarbonyl-marcfortine A.) 'H NMR (300 MHz, CDCI 3 8 0.81 3H), 1.21 3H), 1.19-1.94 8H), 1.41 3H), 1.45 3H), 1.98 1H), 2.08-2.28 2H), 2.40 1H), 2.62 2H), 2.82 1H), 2.99 3H), WO 93/10120 WO 9310120PCT/US92/09483 3.50-3.86 (in, 8H), 4.86 1H), 6.41 1H), 6.85 2H). FAB-MS 591 HRMS M/Z H, C 33
H-
4
N
4 0 6
+H
1 cale. 591.3182, obsd. 591.3200.
Example 18 Following the general procedure of Example 12 but employing the appropriate paraherquamide starting material, there are prepared the following other paraherquamide compounds: I -diethoxyphosphoryl-paraherquamide 1 -cyclopropylcarbonyl-paraherquamide 1 -piperidinyl)thiocarbonyl-paraberquamide 1 -succinoyl-paraherquainide I -(4-carbethoxy-1I,3-thiazolidinin-3-yl)carbony-paraherquaxnide I -phenoxycarbonyl-paraherquaxnide 1 -pyrrolidinylcarbonyl-paraherquamnide I -piperidinylcarbonyl-paraherquamide 1 -(4-morpholinecarbonyl)-paraherquamide Example 19 Following the general procedure of Example 13 but employing the appropriate paraherquainide starting material, there are prepared the following other paraherquaniide compounds: 1 -t-butyryloxymethyl-paraherquamnide 1 -benzoyloxymethyl-paraherqualide 1-acetoxymethy..1 8a-paraherquamide I -3,4-dichlorobenzoyloxy-paraherquarmide Following the general procedure of Example 14 but employing the appropriate paraherquainide starting material there are prepared the following other paraherquainide compounds: 1-(2,4-dinitrobenzenesulfenyl)-paraherquamnide 1-(4-morpholinosulfenyl)-paraherquamnide 1-trichloromethylsulfenyl- 18a-ethyl-paraherquamide I -ethoxycarbhonylsulfenyl- 18a-methyl-paraherquamnide I -palm i toyl-paraherquanide Procedure A: A. Seed Fermentation Cunninghamella echinulata subsp. elegans NRRL 1368, Cunninghamella WO 93/10120 PCT/US92/09483 -46blakesleeana ATCC 8688a, Cunninghamella echinulata subsp. elegans NRRL 1393, Cunninghamella echinulata NRRL 3655 Frozen agar plugs of NRRL 1368 or 1393, ATCC 8688a or preferably NRRL 3655 (stored over liquid nitrogen are aseptically transferred into a 100 ml vol of GS-7 (sterile) medium contained in a 500 ml large mouth shake flask. GS-7 is composed of Cerelose and Pharmamedia, each added at 25 g/l of tap water. The pH of the medium is adjusted to pH 7.2 using ammonium hydroxide. The medium is sterilized by autoclaving for 30 min. Inoculated GS-7 is shaken at 125 to 300 (preferably 250 rpm) at 21 0 -30 0 C, preferably about 28" C, for 24-72 hrs (preferably about 48-72 hours). This fermentation is used to inoculate the biotransformation process at about 1 to about 5% rate (preferably about 5% rate).
B. Biotransformation Process 100 ml vols of GS-7 contained in 500 ml large mouth fermentation flasks are inoculated with the 48-72 hr seed fermentation at about 1 to about 5% rate (preferably about 5% rate).
The inoculated GS-7 is shaken at about 250 rpm at 28 0 C for 24-48 hrs. At this time Marcfortine A, dissolved in dimethylformamide (DMF), is added to the fermentation in a range between 10-12 mg/flask, preferably at 10 mg per 0.4 ml DMF per 100 ml flask/fermentation.
The fermentations containing Marcfortine A are continued as previously indicated for 24-72 hrs.
The contents of 100 flasks, each containing 100 ml fennentation as described in the protocol for biotransformation and which had been shaken at 28 0 C for 24-72 hours following addition of Marcfortine A (0.010 g per flask, 1 g total) in DMF (0.2 ml per flask, 20 ml total) are each treated with CH 2
CI
2 (100 ml). The contents are combined and stirred in a waring blender for 5 minutes. The mixture is centrifuged and the resulting aqueous layer is decanted.
The organic layer is collected, dried (sodium sulfate), filtered and concentrated. The components of the crude mixture are separated by a series of column chromatographies using a silica gel solid extract ratio of 50-125:1. Columns are eluted with either 2-5% methanol in
CH
2 CI, or 15-60% acetone in CH 2
C
2 From these chomatographic separations there is isolated Marcfortine B. HRMS (FAB); m/e, found, 464.2563; calculated for C 27
H
33
N
3 0 4 HI, 464.2549.
If desired, marcfortine B may be crystallized from methylene chloride/acetone. For example, a methylene chloride solution of marcfortine B is concentrated under partial vacuum on a rotatory evaporator. The concentrate is diluted with portions of acetone during the concentration until a slurry of marcfortine B in acetone remains.
SUBSTITUTE SHEET W WA TABLE I Colonia Morphologies of UC 7780 I Medium. Surface ISoluble CY ,A,24 0 C 22-25 mrm umbonate velutinous white to pale entire regular reddish beige to salmon occasional clear sulcate green wiage red wlage pink exudate MEA,24 0 C 25-28 mmn flat floccose, green wiwbite irregular none beige periphery G25N,24 0 C 12-15 mm. umbonate velutinous white wlgreen regular none beige periphery 0 C no germination CYA,37 0 5-10 mm umbonate velutinous white irregular none beige granular WO 93/10120 PTU9/98 PCF/US92/09483 -48- TABLE 11 I ~ANTF1EL!fNE~ C JIRI) PHASE 2 SCREEN 1 Red HC Red TC CPD 4 Ds#Anml urviv.
2 1 MG/JIRD 3 3 89 100 0.33 MG/JIRD 3 3 23 961 0.11 MG/JIRD 3 3 0 0 2A 0.33 MGIJIRD 3 3 76 98 0.11 MG/JIRD 3 3 29 72 12 1 MG/JIRD 3 3 100 100 0.33 MG/JIRD 3 3 99 100 0.11 MG/JIRD 3 3 73 0 0.037 MG/IRD 3 3 12 0.11 MG/JJIRD 2 2 65 61 0.037 MG/JIRD 3 3 8 0 12A 1 MO/JIRD 3 3 99 0.33 MG/JIRD 3 3 28 3 12B 1 MG/JIRD 3 3 100 100 0.33 MG/JIRD 3 3 55 I MG/JIRD 0.31G/RD 100 0.11 IMG/JIDf 3 UL3S TUTE SHEET WO 93/10120 WO 9310120PCr/US92/09483 ANTHELMINTIC JIRD PHASE 2 SCREEN Red HC Red TC CPD Dose #Animals Surviv.
121) 1 MG/JIRD 3 3 94 99 0.33 MG/JIRD 3 3 51 12E 1 MG/JIRD 3 3 99 100 1 0.33 MG/JIRD 3 3 100 100 0.11 MG/JIRD 3 3 80 99 0.037 MG/JIRD 3 2 0 42 12F 1 MG/JIRD 3, 3 99 100 0.33 MG/JIRD 3 3 98 100 r 0. 11 MG/JlRD, 3 3 74 78 120 1 MG/JIRD 3 3 99 100, 0.33 MG/JIRD 3 3 95 100 0.11 MG/JIR 3 3 0 61 12H 0.33 MG/JIRD 3 3 99 99 0.11 MG/JIRD 3 3 2 61 121 1 MG/JIRD 3 3 100 100 0.33 MG/RRD 3 3 100 0.11 MG/JIRD 3 2 69 87 12i 1 MG111RD 3 3 100 100 0.33 MG/JIRD 3 3 96 96 0.11 MG/JIRD 3 3 32 991 0.037 1MO/JIR 3 3 22 44 1 SUBSTITUTE SHEET WO 93/10120 WO 9310120PCT/US92/ 09483 ANTHELNMNTC JIRD PHASE 2 SCREEN Red HC Red TC CPD Dose Animals Surviv.
12K I MG/JIRD 3 3 100 100 0.33 MG/JIRD 3 3 66 54 12L 0.33 MG/JIRD 3 3 99 99 0.11 MGIJIRD 3 3 37 56 12M 0.33 MG/JTRD 3 3 97 100 0.11 MG/JIRD 3 3 73 98 0.11 MG/JIRD 2 2 25 0.037 MG/j'IRD 3 3 11 66 12N 0.33 MG/JIRD 3 3 100 100 F 0.11I MG/JIRD 3 3 81 98, 0.11 MG/JER.D 2 2 0 39 0,037 MG/JlRD 3 3 62 120 0.33 MG/JIRD 3 3 100 100 0.11 MG/IIRD 3 3 80 98 0.11 MG/JIRD 2 2 46 77 0.037 MG/JLRI) 3 3 39 46 12P 0.33 MG/JIRD 3 3 100 100 0.11 MG/JIRD 3 3 51 78 SUBSTITUTE SHEET WVO 93/10120 PCT/US92/09483 ANTHELNTfC JIRD PHASE 2 SCREEN Red HC Red TC CPD Dose Animals Surviv.
12Q 0.33 MG/J1RD 3 3 99 99 0.11 MG/JIRD 3 3 53 63 12R 0.33 MG/JIRD 3 3 100 99 0.11 MG/JIR.D 3 2 96 97 0.11 MG/JIRD 3 3 0 0.037 MO/JIRD 3 3 0.33 MO/JIRD 3 3 99 100 0.11 MG/JJRD 3 3 76 12S 0.33 MG/JIRI) 3 3 100 100 0.11 PMCI/ RD 3 3 93 100.
0.11 MG/JLRD 3 3 29 0.037 MG/JRD 3 3 27 0.33 MG/JIRD 3 3 98 100 0.11 MG/JIRD 3 3 61 97 12T 0.33 MG/JIRD 3 3 74 93 0.11 MG/JIRD 3 3 32 74 12U 0.33 MG/JTRD 3 3 100 99 0.11 MG/IRD 3 3 72 82 12V 0.33 MG/flRD 3 3 98 98 0.11 MGJTRD 3 3 87 86 13 1 MG/fIRD 3 3 100 100 0.33 MG/JRD 3 3 55 SUBSITUTE S EET WO 93/10120 WO 9310120PCf/US92/09483 ANTHELMINTIC JIRD PHASE 2 SCREEN CPD k Dose Animal Surviv. eHC RdT 14 1 MG/JIRD 31 3 100 99 0313 MO/JIRD 3 3 100 100 0.11 MG/JIRD 3 3 74 91 0.037 MG/JTRD 3 3 29 0 0.33 MG/JIIRD 3 3 99 100 14A 0.33 MG/JIRD 3 3 96 100 0.11 MG/JTRD 3 3 81 63 14B I MG/JIRD 3 3 1A 100 0.33 MG/fRRD 3 3 98 100 0.11 MG/JIRD 3 3 0 76, 14C I MG/JIRD 3 2 100 99 0.33 MG/JIRD 3 3 98 100 0.11 MG/JIRD 3 3 29 0.037 MO/JIRD 3 3 56 0 14D I MG/IIRD 3 3 100 100 0.33 MG/JIRD 3 3 100 100 0.11 MG/TIRD 3 3 32 67 MGIJID 3 99 100 0.33 1MG/JIRD 3 3 61_ 76 SUBSTITUTE SHEET WO 93/10120 W093/0120PCf/US92/09483 ANTHELI-NTIC JIRD PHASE 2 SCREEN Red HC Red TC CPD Dose Animals Surviv.
16 1 MG/JlRD 3 3 99 100 0.33 MG/J1RD 3 3 92 100 0.11 MG/JIRD 3 3 50 12 L0,33 Mj/JIRD 1 3 1 3 1 98 100 0.11 MG/JIRD 3 3 76 99 SUBSTITUTE SH1EET WO 93/10120 WO 9310120PCT/US92/ 09483
FORMULAE
H3CCH C 3 N 0 11 N NH
CCR
3 1130 CH3 C 0
NM
5' H
NH
s 0 Compound, 2 Compound 2A Compound 12 Compound 12A WO 93/10120 WO 9310120PCT/US92/09483 Compound 12B Compound 12C Compound 13 WO 93/10120 WO 9310120PCr/US92/09483 Compound 14 Compound 12D Compound C=0
(N
0C WO 93/10120 WO 9310120PCr/US92/09483 Compound 12E 3u 0 N NSc1 0 CR 3 0 CI'/-CJ.
Compound 14B Compound 14D Compound 12F WO093/10120 PTU9/98 PCT/US92/09483 Compound 12G Compound 121 Compound 12J
N
0113 WO 93/10120 PTU9/98 PCT/US92/09483 Compound 12K Compound 14C N v LN 1 0 1SC-OCH3 0 Gil 3 0 Compound 14A WO 93/10120 WO 9310120PCT/US92/09483 Compound 12L Compound 12M Compound 12N WO 93/10120 WO 930120PMU/IY92/09483 Compound 12P 0 Compound 12Q Compound 12R WO 93/10120 WO 9310120PCr/US92/09483 Compound 12S c ,I cl -c-CH Al Compound 12U Compound 12V WO 93/10120 WO 9310120PCr/US92/09483 Compound 12H Compound 120 Compound 12T WO 93/10120 WO 9310120PCT/US92/09483 Compound 16 Marcfortine D
CH
3
NN
0 HC3 0 1C13

Claims (5)

1. A compound of Formula I fl 3 C ,C1 3 R 0O 25, R 2 A CR 3 61 2 !'R N 1 'i.~2NR 1 1 4 1 3 N /N R Rijia 1b1 wherein: n is 0 or 1; R 1 4 and R14b, being the same or different, are selected from hydrogen, hydroxy, C,-C 6 alkyl, C 1 -C 6 alkoxy, Cj-C6 alkenyl, Cl-CG6 alkenyl-Cl-C 6 alkoxy, C 1 -C 6 alkynYl-C 1 -C 6 alkoxy, C,-C6 alkynoyloxy, Poly CI-C 6 alkoxy-C 1 -C 6 alkoxy, phenyl, phenyl-Cl 1 -C6 alkyl, tri-C 1 -C 6 alkylsilyloxy, diphenylphosphoryloxy and halogen, or and R1 4 b together formn the epoxide or with the provisio that when n is 1, then RW 4 and R1 4 b are both hydrogen; m is 0 or 1; W is 0 or S; when W is S, R, is hydrogen, C,-C7 alkyl, cyclo C 3 -C~alkyI, benzyl, C.2-C7 alkanoyl (-C(O)C 2 -Catkyl) (optionally substituted with carboxy (-COON), C,-C7 alkanoyl, carbo C,-C~alkoxy (-C(O)OCI-Calkyl), -NR 4 R 5 axninocarbonyl (-C(O)NR 4 R 5 I, C 10 -C24atlcanoyl (-C(O)Cl-C2 4 alkyl, cyclo C 3 -C 8 alkanoyl (optionally substituted with carboxy, C,-C 7 alkanoyl, carboo 1 -Cqalkoxy, -NR 4 R 5 aminocarbonyl), alkanoyloxymethylene (-CH 2 OC(O)-C2-Calhyl), benzoyloxymethlene (-CH- 2 0C(O)phenyl) [optionally substituted with 1 or 2 groups selected from halogen, C 1 -C 4 alkyl, haloCj-C~alkyl, nitro, cyano and C 1 -C~alkoxy), C 10 -C24alkenoyl (-C(O)C 9 -C2 3 alkenyl), benzenesulfonyl (-SO 2 CH 2 phenyl) (optionally substituted with I or 2 groups selected from halogen, Cj-C4 alkyl, haloC,-Calkyl, nitro, cyano and Cl-C7alkoxyl, C 1 -C 4 alkylaininocarbonyl (-C(O)N(C,-Cr-XV2) 2 C 1 -C 4 alkylam inothiocarbonyl (-C(S)N (C 1 -C: 4 alkyl) 2 Cj-C7 alkoxycarbonyl, phenoxycarbonyl [optionally substituted with 1 or 2 groups selected from halogen, C,-C 4 alkyl, haloC,-C~alkyl, nitro, cyano and C 1 alkoxy), -C(O)NR' 4 R' 5 3 -SR6, -SO 2 NR 4 R5, benzoyl substituted at the 3 or 4 position with WO 93/10120 WO 9310120PCJJ/US92/09483 -66- -CH 2 NR 4 R 5
2-tetrahydrofuran, or bicycoC 8 -C, 2 alkanoyl; when W is 0, R, is selected from the group consisting of: C 2 -C 7 alkanoyl substituted with carboxy C 1 -C 7 alkanoyl, carboC,-C 7 alkoxy (-C(0)0CI-C 7 alkyl), -NR 4 R 5 aminocarbonyl (-C(O)NR 4 R 5 cyclo C
3 -C~alkanoyl optionally substituted with carboxy, C 1 -C 7 alkanoyl, carboCI-C~alkoxy, -NR
4 R 5 aminocarbonyl; alkanoyloxymethylene (-CHOC(O)-C 2 -C 7 alkyl); benzoyloxymethlene (-CH 2 OC(O)phenyl) substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, haloC,-C 7 alkyl, nitro, cyano and Cl-Cqalkoxy; phenoxycarbonyl substituted with I or 2 groups selected from halogen, CI-C 4 alkyl, haloCI-C 7 alkyl, nitro, cyano and C,-C 7 alkoxy; (M -C(0)NR' 4 R' 5 2 )(R 3 -SR 6 C 1 -q 4 alkenoyl (-C(0)C 9 -C 2 3 alkenyl); or 2-tetrahydrofuran; R 4 and being the same or different, are selected from hydrogen, Cj-C 7 alkyl, phenyl (optionally substiwted with I or 2 groups selected from halogen, CI-C 4 alkyl, haloCI-C 7 alkyI, nitro, cyano and C,-C 7 alkoxyl or when taken together with N, form a saturated or unsaturated heterocyclic amine ring; R' 4 and R' 5 being the same or different, are selected from C 1 -C 7 alkyl, cyclo(C3-Cs)alkyl, phenyl 'optionally substituted with I or 2 groups selected from halogen, C,-C 4 alkyl, ha]oC,-C 7 alkyl, nitro, cyano and C,-C 7 allcoxy) or when taken together with N, form a saturated heterocyclic amnine ring optionally containing I or 2 additional heter-oatoms selected from N, 0 or S; X is 0 or S; R 2 and R 3 being the same or different, are selected from CI-C 7 alkyl, phenyl [optionally substituted with I or 2 groups selected from halogen, C 1 -C 4 "Ikyl, haloC 1 -C~allcyl, nitro, cyano and CI-C 7 alkoxy), C,-C 7 alkoxy, thio(C,-C 7 )alkoxy, phenoxy, thiophenoxy, -NR 7 R8 (where R 7 and R 8 being the same or different, are selectd from H, CI-q7 alkyl or taken together with N, form a saturated heterocyclic ring), or taken together with P form a 4- to 7-membered heterocyclic ring; RO s C,-C 7 alkyl, haloC,-C 7 alkyl, carboC 1 -C 7 alkoxy, -NRR 1 where and Rio, being
4585.PCP -67- the same or different, are C,-C alkyl or phenyl (optionally substituted with I or 2 groups selected from halo, lower alkyl, haloC 1 -Cqakyl, nitro, cyano, CI-Cq alkoxy); R. is hydrogen, halogen or C,-C 7 alkoxy; is hydrogen or halogen; is hydrogen, CI-C alkyl, C 2 -Cs alkoxyalkyl, C 2 -Cs alkenyl, C2-C 1 alkynyl or benzyl; the broken line between carbons 24 and 25 represents a single or double bond; and pharmaceutically acceptable salts and hydrates thereof; with the overall proviso that, when W is 0, RI4 and R' 5 are not both C1-C 7 alkyl. 2. A compouid maccordin lamIweei is 0. 3. A compound according to claim 2 where R, and R5 are hydrogen; R18. is hydrogen,- C,-C 7 alkyl, C,-Cs alkoxymethyl, C 2 -qC alkenyl or benzyl; and the broken line represents'a double bond between carbons 24 and 4. A compound according to claim I where W is S and n is 0. A compound according to claim 4 where m is 0; R. and Rz are hydrogen; R 1 U is hydrogen, C 1 -C 7 alkyl, C,-Cs alkoxymethyl, Ca-C, alkenyl or benzyl; and the broken line represents a double bond between carbons 24 and 6. A compound according to claim 5 which is selected from 18-thiomarcfortine A; 1-acetoxymethyl-I 8-thiomarcfotine A; "ietoxyphosphozl- 1 8-thiomarcfortine A; 1 -dimethylsulfamoyl- 18-thiomnarcfortine A; 1-cyclopropylcarbonyl-I8-iomarcfortine A; 1-(1 -piperidinyl)thiocarbonyl- 18-thiomarefortine A; I-succinoyl-1 8-thiomarcfortine A; 1-(2,4-dinitrobenzenesulfenyl)-18-thomarcfortine A; 1 -(4-morpholinosulfenyl)-1 8-thiomarcfortine A; 24-propoxy-24,25-dihydro- I 8-thiomarcfortine A; 1 -(p-toluenesulfonyl)- 18-thiomarcfortine A; i-acetyl-18-thiomarcfordne A; 1 1-methyl-18-thiomarcfortine A; l-benzyl-18-thiomarcfortine A; SUBSTITUTE SHET WO 93/10120 WO 9310120PCr/US92/09483 -68- 1-dimethylcarbamoyl-1 8-thiomarcforie A; 1-methoxycarbonyl-18-thiomarcfortine A; I 8-thiomarcfortine B; 24,25-dihyciro-1I8-thiomarcfortine B; 24-niethoxy-24,25-dihydro-1I8-thiomarcfortine B; 1-(p-toluenesulfonyl)- 18-thiomarcfortine B; 1-acetyl-1I8-thiomarcfortine B; I1-ethyl- I 8-thiomarcfortine B; I -benzyl-1 8-thiomarcfortine B; 18a-ethyl-18-thiomarcfortine B; I 8a-benzyl-1I8-thiomarcfortine B; I 8a-methoxyethoxymethyl-18-thiomarcfortine B; 18a-allyl-18-thiomarcfortine B; 1 8a-propargyl-18-thiomarcfortine B; 1 8a-ethyl-24-methoxy-24,25-dihydro-1I8-thiomarcfortine B; 18 ethyl-1I8-benzyl- I8-thiomarcfortine B; I 8a-ethyl-24-methoxy-1 8-thiomarcfortine B; 14-2,4-dinitrobenzene sulfonyl)- 18a-ethyl-24-methoxy-24,25-dihydro- 18-thiomarcfortine B; 18a-ethyl-24,25 dihydro-18-thiomarcfortine B; 1 8-thiomarcfortine C; 24,25 dihydro-18-thiomarcfortine C; I -(p-bromobenzene sulfonyl)- 18-thiomarcfortine C; I -propionyl-1 8-thiomarefortine C; 1-propyl-18-thiomarcfortine C; 1-benzyl-1 8-thiomarcfortine C; 1 8a-propyl- 18-thiomarcfortine C; I 8a-benzyl- 18-thiomarcfortine C; 18a-nriethoxyethoxynetihyl-1 8-thiomarofortine C; 18a-allyI-18-thiomarocfortine C; 1 Sa-propargyl- 18-propyl- 18-benzyl- 1 -thiomarcfortine C; 1 -(4-carbethoxy- l,3-thiazolidinin-3-yl)carbonyl-1 8-thiomarcfortine A; 1-palmnitoyl-18-thiomarcforitine A; or I -(4-morpholinocarbonyl)- 1 -thiomarcfortine A. 7. A compound according to claim 1 where W is S and n is 1. WO 93/10120 WO 9310120PCT/US92/09483 -69- 8. A compound according to claim 7 where m is 0; R24and R.5 are hydrogen; RI, is hydrogen, C,-C 7 alkyl, C,-Cg alkoxymethyl, C 2 alkenyl or benzyl*; and the broken line represents a double bond between carbons 24 and 9. A compound according to Claim 8 where R 14 and RJOb are both hydrogen. A compound according to Claim 8 where R1 4 is methyl, R1 4 b is hydroxy. 11. A compound according to claim 8 which is selected from 1 8-thoparaherquamide 24,25-dihydro-1 8-thioparaherquamide 14-0-methyl- I 8-thioparaherquamide 14-0-ethyl- I 8-thioparaherquamide 14-0-butyl- 1 8-thioparaherquamide 14-O-benzyl-1I 8-thioparaherquaxnide 14-0-ally- I 8-thioparaherquainide 14-0-propargyl- I 8-thioparalierquamide 14-0-methoxymethyl- 1 8-thioparaherquaniide 14-0-methoxy-ethoxy-1 8-thioparaherquamnide 14-0-methoxy-ethyoxy-methyl-18-thioparaherquamide 17-methyl- I 8-thioparaherquamide 17-methyl ene- I 8-thioparaherquamide I -N-(p-toluenesulfonyl)-1I 8-thioparaherquamnide 24-methoxy-24,25-dihydro- I 8-thioparaherquaxnide 12. A compound according to Claim 1 wherein W is 0 and n is 0. 13. A compound according to claim 12 where m is 0; R24 and R 2 5 are hydrogen; RI,, is hydrogen, C 1 -C 7 alkyl, C 1 -Cg alkoxymethyl, C 2 -C 8 alkeiiyl or benzyl; and the broken line represents a double bond between carbons 24 and 14. A compound according to claim 1 which is selected from I -piperidinyl)thiocirbonyl-marcfortine C; 1 -piperidinyl)thiocarbonyl-marcfortine A; 1 -(2,4-dichlorophenoxy)carbonyl-1I8a-N-ethyl-24-methoxy-24,25-dihydro-marcfordne B; 1 -(2,4-dichlorophenoxycarbonyl)- 18a-allyl-marcfortine B; WO 93/10120 WO 9310120PCr/US92/09483 1 -(2,4-dinitrobenzenesulfenyl)-marcfortine A; 1-(3-acetoxy)propionyl- 18a-N-propargyl-marcfortine B; 1 -(4-carbethoxy- 1,3-thiazolidine-3-yl)carbonyl-marcfortine A; 1 -(4-morpholinosulfenyl)-marcfortine C; 1-(4-oxobutyryl)- 18a-methyl-marcfortihe C; 1-(4-oxopentanoyl)-marcfortine A; I -(4-oxopentanoyl)-marcfortine C; 1-3,4-dichlorobenzoyloxy-rnarcfortine C; 1 -acetoxym ethyl marcfortine C; 1 -acetoxyrnethyl- 18a-benzyl-marcfortine B; 1 -benzoyloxymethyl-marcfortine A; 1 -cyclohexylcarbonyl-niarcfortine A; 1-cyclohexylcarbonyl-marcfortine C; I -cyclopropylcarbonyl-marcfortine A; 1 -cyclopropylcarbonyl- 18a-ethyl-marcfortine B; 1-cyclopropylcarbonyl-marofortine C; 1-diethoxyphosphoryl-marcfortine A; 1-diethoxythiophosphoryl-marcfortine A; 1 -diethoxythiophosphoryl-marcfortine C; 1 -dimethylamninoacetyl-marcfortine C; 1 -dimethylaminoacetyl-marcfortine A; 1-dirnethylamninosulfenyl- 18a-N-benzyl-marc'fortine B; 1 -diphenylp~hosphinyl-1I a-N-aIlyI-marcfortine B; 1-ethoxycarbonylsulfenyl-1 8a-methyl-marcfortine C; 1 -ethoxycarbonylsulfenyl- 18a-N-methoxyethoxym ethyl-marcfortine B; I -N-(2,4-dinitrobenzenesulfenyl)-1 8a-N-ethyl-marcfortine B; 1-N-(4-carbethoxy- 1,3-thiiazolidinin-3-yI)carbonyl-marcfortine C; 1 -N-(4-carbethoxy-1,3-thiazolidinin-3-yl)carbonyl-marcfortine A; 1 -N-(p-bromobenzenesulfenyl)-inarcfortine C; 1-N,N-dimethylsuccinamnidoyl-inarcfortine A; 1 -N,N-dimethylsuccinamidoyl-marcfortine C; 1-phenoxycarbonyl-marcfortine C; 1-phenylmethoxyphosphonyl-rnarcfortine A; 1 -phenylmethoxyphosphonyl-marcfortine C; 1-succinoyl-marcfortine A; I -succinoyl-marcfortine C; WO 93/10120 WO 93/10120PCT/US92/09483 -71- 1-t-butyr-yloxymethyl-marcfortine A; or I -trichloromethylsulfenyl- 18a-ethyl-marcfortine B. A compound according to Claim I selected from 1 -cyclopropylcarbonyl-marcfortine A I -phenoxycarbonyl-marcfortine A 1 -palm itoyl-marcfortine A I -[(4-nitrophenyl)oxylcarbonyl]-marefortine A 1 -pipeiidinecarbonyl)-marcfortine A 1-f [4-(ethoxycarbonyl)piperazin-1I-yR jcarbonyl]-marcfortine A 1-[14-(benzyl)piperazin- 1-yllcarborij-marcfortine A 14 ,3-benzodioxol-5-yl-methyl)piperazin-1I-yllcarbonyl J-marcfortine A 1 J4-(methyl)piperazin-1I-ylllcarbonylll-marcfortine A 1 ,3-dihydro- 1,3-dioxo-2H-isoindol-2-yl)-marcfortine A 1 [4-(pyridin-2-yI)piperazin-1 -yljlcarbonyl)-marefortine A I -Ii 4-(phenyl)piperazin 1 -ylllcarbonyN]-marcfortine A I -[[4-(chlorocarbonyl)piperazin-1-yljcarbonyl]-marcfortine A 1-[[4-(phenyl)piperidin-1-yllcarbonyl]-marcfortine A I t4-(phenyl)(dimethyl)piperidin-1I-yllcarbonylj-marcfortine A 1 [4-(5..choropyridazin-3-yl)piperazin-1I-yllcarbonylJ-marcfortine A 1 [4-(4-chlorophenyl)piperazin-1I-ylilcarbonylll-marcfortine A I [4-(trichloroethoxycarbonyl)piperazin-1I-yl]carbonyl ]-ma rcfortine A I -[i[4-(3-trifluoromethyl-thiadiazol-I -yl)piperazin- 1-yljcarbonyl]-marcfortine-A I -acetoxymethyl-marcfortine A 1 -(2,4-dinitrobenzenesulfenyl)-marfortine A I -(4-morpholinosulfenyl)-niarcfortine A 1 -(trichloromethylsulfenyl)-xnarcfortine A 1 -(methoxycarbonylsulfenyl)-inarcfortine A I -(benzenesulfenyl)-marcfortine A 1 -(2-tetrahydrofuranyl)-marcfortine A, l-(4-rnorpholinocarbonyl)-marcfortine A or I -(4-morpholinocarbonyl)-marcforine A N-oxide. 16. A compound according to claim 1 selected from 1-diethoxyphosphoryl-paraherquamnide 1-cyclopropytcarbonyl-paraherquamnide WO093/10120 PTU9/98 PCr/US92/09483 1 -piperidinyl)tdocarbony-parahierquamide I-succinoyl-paraherquarmide I -(4-carbethoxy-I ,3-thiazolidinin-3-yl)carbonyl-paraherquamide I -phenoxycarbonyl-paraherquamide 1 -pyrrolidinylcarbonyl-paraherquamide I -t-butyryloxymethyl-paraherquamide I 'benzoyloxymethyl-paraherquamide I -acetoxymethyl-1I 8a-paraherquamide I -3,4-dichlorobenzoyloxy-paraherquamide 14-2,4-dini trobenzenesulfenyl)-paraherquamide I -(4-rnorpholinosulfenyl)-paraherquamide I '-trichloromethylsulfenyl- I 8a-ethyl-paraherquamide I -ethoxycarbonyisu Ifenyl- I 8a-methyJ-parherquamide I -palmnitoyl-paraherquamide I-piperidinylcarbonyl-paraherquamid1e 1-(4-morpholinecarbonyl)-para!~erquamide. 17. A method for the trearin t or prevention of helminth or arthropod infections in domesticated animals which c prises treating such animals with an effective am nt of a compound of claim 1. 18. A method for e treatment of insect or nematode pests plants which comprises treating said plants r the soil in which they grow with iective amount of a compound of claim 1. 2519. A omposition useful for the tr ent and prevention of helm inth or arthropod infecti s of domesticated anima hich is comprised of an inert carrier and a compound of cla 1,. 20. A ccom sition useful for the prevention and treatment of insect or nematode pests of plants w ih n is comprised of an inert carrier and a compound of claim 1. 73 17. A compound according to any one of the preceding claims, wherein R, is C(O)NR' 4 R' 5 and NR' 4 R' 5 is a saturated heterocyclic ring optionally containing I or 2 additional heteroatoms selected from N, 0 and S. 18. A compound according to claim 17, wherein the saturated heterocyclic ring is selected from 4-norpholine 4-phenyl-i -piperazine 4-(2-pyridinyl)-l -piperazine 2,6-dimethyl-4-morpholine 1-pyrrolidine 4-methyl-I -piperazine I -piperidine 4-phenyl-1 -piperidine thiazolidine *:is 4-pheny-1 ,2,3,6-tetrahydropyridine 4-phenylpiperidine .ethyl prolinate tetra hyd rofu rylamine 3-pyrroline thiazolidine-4-carboxylic acid thiomorpholine nipecotamide 2-methylpiperidine o 3-methylpiperidine 4-methylpiperidine N-methylpiperidine I -methyihornopiperazine I -acetylpiperazine and N-carboethoxypiperazine. 19. A method for the manufacture of a medicarnent for use in the treatment or prevention of helminth or arthropod infections in domesticated mammals including the step of bringing a compound of any one of the preceding claims into a form suitable for 7~ R~iLadministration. C)WNC'IWZNWOWWEDY)TYPING)31264E.DC 74- A method for the treatment of insect or nematode plant pests, which includes applying a compound according to any one of claims 1 to 18 to the plants or the soil in which they grow. 21. A biologically pure culture of Penicillium sp. NRRL 18887. c- Nr woOewantrtYmNG24T.Doc it~ C WYO 93/10120 PCT/US92/09483 22. A process for producing a compound of the formula: 6 0 7 24 which compises- the biotransformation of a compound of the formula: H 3 6 16 H 3 C CH 3 16 123 3 13 3 1 N 14 '19N 1R N using a microorganism in an aqueous nutrient medium containing an assimilable source of carbon and an assimilable source of nitrogen under aerobic conditions. 23. The process of Claim 22 wherein the i icroorganism is a species of Cunninghamella or a mutant thereof. 24. The process of Claim 22 wherein the strain is selected from the group consisting of NRRL 1368, NRRL 1393, NRRL 3655 or ATCC 8688a. WO 93/10120 WO 9310120PCT/US92/09483 A compound selected from the group consisting of 10"-(1I,4-dicarbonylpiperazine)bis(6',7',8',9', 10', 1 0'a-hexahydro- 1 ',4,,12'-pentamethyl) alpha., 3'aalpha. ,9'a.alpha., 1 aS*, 1 I 0'a.beta.1] -Spiro [4H,8H-[ 1 ,4]dioxepino [2,3-glindole-8,2'(3'H)- [1 H,4H- 3a,9a7(iminomethano) cyclopentab]quinolizin]-9,1 1'(10H)-dione; or 1,4-dioxo-2-butene)bis(6',7', 10', 10'a-hexahydro- 1 12'-pentarriethyl)-[2' S-[2'.alpha. ,3'a.alpha. ,9'a.alpha., 1 -Spiro[4H,8H-[ 1 ,4]dioxepino[2,3-g]indole-8,2'(3'H)-[ 1 H,4H-3a,9,fl (iminomethano)cyclopentalb] quinolizinl-9, I1'(1 OH)-dione. I" LL INTERNATIONA sEARCH REPORT Intlernaonal Application~ No PCT/US 92/09483 I. CLASSIFICATION0OFSUBJECTMA7rER (if seveual classification symbols apply, indicato all) 6 Int.Cl.
5 C07D491/22; A01N43/90; C12P17/18; C12NI/14 011W AI;1 1 r i. d(C7D49/22,321:00,241:00, 221:00, 221:00, 2 09:00) Documentation Saarched other than Minimum Documentation to the Extent that such Documents are Included In the Fields Searched 1 i11. DOCUMENTS CONSIDERED TO BE RELEVANT 9 category Citation of Document, It with indication, where appropriate, of the relevant passages 12 Relevant to Claim No.1 3 X US,A,4 923 867 A. BLIZZARD ET AL.) 1,6,14, 8 May 1990 19,20 cited in the application see claims 1,18,19 X EP,A,O 354 615 (MERCK CO) 1,19,20 14 February 1990 cited in the application see claims 1,9,10 X TETRAHEDRON LETTERS. 1,19,20 vol 32, no. 22, 1991, OXFORD GB pages 2441 2444 T. A. BLIZZARD ET AL. 'Chemical modification of paraherqTuamide. 4. 1-N-Substituted analogs 1 see the whole document oSpecial categories of cited documents: :10 IT' later document published after the International filing date A doumet dfinng he eneal sateof he rt hic Isnotor priority date and not In conflict with the application but ''dcundein he eart l rSteae h nwihIo cited to understand the principle or theory underiying the consdere tobe o paticuar eleniceinvention earlier document but published on or after the International IX' document of particular relevance; the claimed Invention filing datc cannot be considered novel or cannot be considered to 1. document which may throw doubts on priority claim(s) or Involve an Inventive step which Is cited to establish the publication date &f another document of particula.- relevance; the claimed Invention citation or other special reason (as speifid) cannot be considered to Involve ain Inventive step when the 'Q document referring to a oral disclosure use, exhibition or document Is combined with one or wm other such docu- other means ments, such combination being obvious to a person skilled *11' document published prior to the International filing date but in the art later than the priority date claimed S1 document member of the Same patent family IV. CERTIFCATION Date of the Actual Completion of the International Starch Date of Mailing of this Interaationai Search Report 16 MARCH 1993 26. 03. 93 International Searching Authority Signature of Authorized Officer EUROPEAN PATENT OFFICE VOYIAZOGLOU D. Fome PCT/15A1210 tsecemd aiwd) (Jvieuy 1993) PCT/US 92/09483 Inteuticrsi Appikation No ill. DOCUMENTS CONSIDERED TO BE RELEVANT (CONTIENUED FROM THE SECOND SHEET) C*vcgor atition of Doazment, with Indication, whome appwpdate 1 of the reloant pasoqes Rd a to aam No. A JOURNAL OF ORGANIC CHEMISTRY.I vol. 54, no. 26 May 1989, EASTON US pages 2657 2663 T. A. BLIZZARD ET AL. 'Chemical modification of paraherquamide. 1. Unusual reactions and absolute stereochemistry' see page 2658, scheme III A TETRAHEDRON LETTERS. 1,22 vol. 22, no. 21, 1981, OXFORD GB pages 1977 1980 T. PRANGC ET AL. 'Structures of marcfortine B and C X-ray analysis alkaloids from Penicillium Roqueforti' cited in the application see the whole document Pau PCrfMAh1o 443ihi&ba) (jWWasy ANNEX TO THE INTERNATIONAL SEARCH REPORT ON INTERNATIONAL PATENT APPLICATION NO. US 9209483 SA 67811 This annex lists the patent family members relating t o the patent documents cited ia the above-aientioned intcrnational search report The members ame as contained in the European Patent Office EDP file on The European Patent Office is in no way liable for these particuar which are merely given for the purpose of information. 16/03/53 Patent document Publicato Patent family Publication cited in search report I dat member~s) Id aft I US-A-4923867 08-05-90 None EP-A-0354615 14-02-90 US-A- 4978656 18-12-90 JP-A- .2149581 08-06-90 C~ For more details about this annex seme official journal of the European Patent Office, No. 12/82
AU31264/93A 1991-11-22 1992-11-13 Marcfortine/paraherquamide derivatives useful as antiparasitic agents Expired - Fee Related AU681998B2 (en)

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
US79545791A 1991-11-22 1991-11-22
US79694891A 1991-11-22 1991-11-22
US79590091A 1991-11-22 1991-11-22
US79773691A 1991-11-22 1991-11-22
US795457 1991-11-22
US795900 1991-11-22
US797736 1991-11-22
PCT/US1992/009483 WO1993010120A1 (en) 1991-11-22 1992-11-13 Marcfortine/paraherquamide derivatives useful as antiparasitic agents
US796948 2004-03-10

Publications (2)

Publication Number Publication Date
AU3126493A AU3126493A (en) 1993-06-15
AU681998B2 true AU681998B2 (en) 1997-09-18

Family

ID=27505792

Family Applications (1)

Application Number Title Priority Date Filing Date
AU31264/93A Expired - Fee Related AU681998B2 (en) 1991-11-22 1992-11-13 Marcfortine/paraherquamide derivatives useful as antiparasitic agents

Country Status (5)

Country Link
EP (1) EP0613479A1 (en)
JP (1) JPH07501693A (en)
AU (1) AU681998B2 (en)
CA (1) CA2121234A1 (en)
WO (1) WO1993010120A1 (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3134936B2 (en) * 1992-10-16 2001-02-13 沖電気工業株式会社 Impact sensor
TW410227B (en) * 1993-06-16 2000-11-01 Upjohn Co 14-substituted marcfortines and derivatives useful as antiparasitic agents
AU4911097A (en) * 1996-11-15 1998-06-03 Pharmacia & Upjohn Company 1- and 2-substituted marcfortines and paraherquamides as antiparasitic agents

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4923867A (en) * 1988-08-24 1990-05-08 Merck & Co., Inc. Synthetic marcfortine derivatives useful as antiparasitic agents
US4978656A (en) * 1988-08-12 1990-12-18 Merck & Co., Inc. Synthetic derivatives of paraherquamide

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4978656A (en) * 1988-08-12 1990-12-18 Merck & Co., Inc. Synthetic derivatives of paraherquamide
US4923867A (en) * 1988-08-24 1990-05-08 Merck & Co., Inc. Synthetic marcfortine derivatives useful as antiparasitic agents

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TET. LETT., 32 (22) PP 2437-2440 (1991) *

Also Published As

Publication number Publication date
AU3126493A (en) 1993-06-15
CA2121234A1 (en) 1993-05-27
EP0613479A1 (en) 1994-09-07
WO1993010120A1 (en) 1993-05-27
JPH07501693A (en) 1995-02-23

Similar Documents

Publication Publication Date Title
EP1489082B1 (en) Intermediate compounds
US5776936A (en) Marcfortine/paraherquamide derivatives useful as antiparasitic agents
GB2166436A (en) Antibiotic compounds and their preparation
US4978656A (en) Synthetic derivatives of paraherquamide
IE42605B1 (en) Rifamycins
US5075307A (en) Paraherquamide and dihydroparaherquamide as antihelminthic agents
AU681998B2 (en) Marcfortine/paraherquamide derivatives useful as antiparasitic agents
US4873247A (en) Derivatives of paraherquamide isolated from a fermentation broth active as antiparasitic agents
US5164389A (en) Anthelmintic bioconversion products
EP0703916B1 (en) 14-substituted marcfortines and derivatives useful as antiparasitic agents
CA1236785A (en) ANTIBIOTIC LL-D42067.beta.
AU647265B2 (en) Anthelmintic milbemycin analogs from streptomyces hygroscopicus strains
RU2107687C1 (en) Derivatives of markfortine/paragerkwamide, method for suppressing of pests, insect and fungi nematodes