MXPA97005562A - Improved synthesis of cyclopropilacyl - Google Patents

Abstract

The present invention relates to cyclopropylacetylene by mixing a strong base and 5-halo-1-pentyne in an aprotic solvent, the temperature is allowed to rise from 0 to 150 ° C and this temperature is maintained for at least 15 minutes, then the mixture of reaction is extinguished with a proton source

Description

IMPROVED SYNTHESIS OF CICLQPRQPILACETILENQ BACKGROUND OF THE INVENTION This case is related to the Case ñorcr-- 18793TB, which is a continuation in part of 18793IA, which is a continuation in part of the Merck Case 18793, presented on (August "Je 1992, U. .SN 07 / 925, 1-107 and Case 19344. A backbone designated as ninunodefic human virus (HIV) is the etiological agent of the complex disease "which includes progressive destruction of the immune system (acquired nrnundodefíciencia syndrome, STDñ ) and degeneration of the central and peripheral nervous system This virus was previously known to be an LAV, HTLV-III or ARV.A common feature of the replication of re + rovius is the reverse transverse transformation of the genome. RNA is used to generate DNA copies of HIV sequences, a step required in viral replication, and it is known that some compounds are reverse loop transcppt inhibitors and are effective agents. in the treatment of ID and similar diseases ares, v.gr.,? gone * i midi na or AZT. The VTH nucleotide sequence shows the presence of a pol gene in an open reading frame [Ratner, L. et al., Nat re, 313, 277 (1985) 3. Homology in the amino acid sequence provides evidence that the sequence of o 1 codic ranscp pt asa mversa, an endonuclease and an HIV protease ["Ton, H. et al., EMBQ 3. ^ 1267 (1985); Powor, RI.D. et al., Science, 231, 1567 (1986), Pearl, LH et al., N tu re, 329, 351 (1987) 1. Applicants demonstrate a substantially more substantial synthesis than that of a HIV reverse transferase inhibitor, the following is ruc ura Denominate swim (-) 6- chloro- 4- ci clo? rop? let? n? l-4- t p luoromet i i -i, 4- < 1? hi dro-2H-3, 1 -ben ^ oxai n ~ 2 -one, hereinafter "Compound A" "You have been found to be highly potent, even against HIV reverse transferase resistant to other SILe antiviral compounds. Applicants have contemplated a substantially improved synthesis of cyclopropylacetylene, an intermediate of Compound A. The foregoing methods used two-step procedures using corrosive reagents, and proceeded in low overall yields. See, for example, mio.tz.er, H.C et al., Synthesis, 998 (1993); Schoberth, W. et al., Synthesis, 703 (1972) CPC15 V base]; Sherrod, and others, J.ñin. Che. oc, 93: 8, 1925-1940 (April, 1971) CI2 on ho drazona]; Mikhailov and Bronovit kaya, Zh. Obshch. Khi Vol..XXII, 195-210 (1952) Cdibromuro]. In contrast, this procedure is shorter than previous methods, does not use corrosive reagents, and produces an overall performance as good or better than that of the previous methods. The present "friction" includes cyclization of 5-halo-1-entmo on strong base. The applicants have discovered that the successful performance of this reaction requires the generation of a transient dianion which is cyclized to cyclopropylacetylene. No method of the technique generates a dianion. On the contrary, the technique indicates that a variety of side reactions may occur, including displacement of chlorine with the base or the deprotonated acetylene, or halogen metal exchange of the odor-uro.

BRIEF DESCRIPTION OF THE INVENTION An improved synthesis of cyclopropyi acetylene, an intermediate of Compound A, is disclosed. Synthesis includes cyclization of 5-halo-1-enti on a strong base. Compound A is useful in the inhibition of HIV reverse transcppt (and its resistant varieties), the prevention of HIV infection, the treatment of HIV infection and in the treatment of AIDS and / or ARC, either as compounds , pharmaceutically acceptable salts (where appropriate), or pharmaceutical ingredient ingested, either in combination or otherwise, with other antimicrobials, immunomodulators, antibiotics or vaccines. Five methods of treating AIDS, methods of preventing HIV infection, and methods of treating HIV infection are also described.

DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED MODALITIES L? The process of the present invention is directed to preparing cyclopropylacetylene, a useful reagent for adding a cyclopropylacetylene group to a large variety of antiviral agents and other medicinal compounds, especially the reverse transcriptase inhibitors of VTH. In this invention, the method of preparing full-length dclopropyl * comprises the steps of (a) mixing at least about 1.0 and "ju? valent is a strong base in atropic solvent with an equivalent of 5-halo-1 -pent in aprotic solvent at a temperature between about -20 and approximately 1 0 ° C; (b) "Let the temperature of the reaction mixture increase to a scale between about 0 and about 150 ° C and keep the temperature within the scale for at least 15 minutes, or until the cyclization is over. be substance Limo complete; and (c) extinguish the reaction with any source of protons. In one embodiment of the present invention, the process of preparing cyclopropylacetylene comprises the steps of (a) mixing at least about 1.0 equivalents of base in aprotic solvent with a 5-halo-1-pentane in Appropriate solvent at a temperature between about -20 and about 150 ° C; (b) Allow the temperature of the reaction mixture to increase to a scale between about 0 and about 150 ° C and keep the temperature within the scale for at least 15 minutes, or until the cyclization is substantially complete; and (c) cooling the reaction mixture to a temperature between about -30 and about 50 ° C; (d) quench the reaction with any proton source. In another embodiment of the present invention, the final step of purifying the desired product of the full-strength cocktail is added. A preferred embodiment of the present invention is a process for the preparation of cyclopropylacetylene comprising the steps of (a) mixing in re about 2.0 and about 2.5 equivalents of n-butyllithium in cyclohexane with an equivalent of 5-chloro-1-pentane in cyclohexane to appro priately f) ° C; (b) heat the reaction to approximately 75 ° C and maintain the reaction at that temperature for approximately 5 hours, or until the cyclization is substantially constant; (o) cooling * the reaction mixture to apr-oximedarnent at 0 ° C; and (cl) quenching the reaction with saturated NH 4 Cl; and optionally, (e) purify the product desired by the cyclopropyl 1 acetyl ene. The process of the present invention is a single-vessel process which begins with the mixing of an equivalent of 5-halo-1-pen ino in aprotic solvent with at least about 1.0 equivalents of strong base in aprotic solvent at a temperature from apr-ox to -20 to approximately 150 ° C. A preferred scale of strong base equivalents is between about 2.0 and about 2.5 equivalents. A preferred starting material is 5-chloro-1 -pentino. A preferred temperature for this mixing is within the range of between about -20 and about 25 ° C, preferably about 0 ° C. Before mixing, the aprotic solvent for 5-h lo-1-μen i may not or may not be the same as the aprotic solvent for the strong base. The strong base is selected from the group consisting of n-butyllithium, sodium amide, sodium ilaride sodium, sodium hydride, potassium hydride, sodium (tr? Methylsilyl) ammonium sodium, b? s (t rimetilsi li Damida potassium, LDA, sec-butill t, o, t-butii lithium, and tetrarnetylpipepide lithium.A preferred strong base is n-butyllithi.The aprotic solvent is selected from THF, 2,5-dimethyl-THF, 1,4-d-oxane, MTBE, diethoxy ethane, di-ethoxyethane, cyclohexane, hexane, and hexane with ethylene diamine, A preferred aprotic solvent is heavenhexane. and 5-halo-1 -pentino is an exothermic reaction that causes cyclization Cyclization occurs spontaneously It is preferred to heat the reaction sufficiently to accelerate cyclization A preferred temperature for cyclization is within the range of about 50 to about 80 ° C, preferably at approximately 75 ° C. The higher the temperature, the shorter the incubation time necessary to substantially complete the cyclization. For a temperature of 75 ° C, an incubation time of about 5 hours is typically required for <;, om? le + ar cyclization. It will be understood that an experienced technician can easily determine temperature variations and incubation time in this cyclization step. When the cyclization is substantially complete, or at least sufficiently complete, the reaction mixture can optionally be cooled to a temperature of between about -30 and about 50 ° C, preferably a temperature of about 0 ° C. Next, a source of protons is added to extinguish the reaction. In this invention, the proton source is selected from saturated NH 4 Cl, HCl and H 2 O *. One source of protons proffered is NH CI. Finally, a purification step can be included at this point to isolate the ecycleproduct of the wood. The reactions used to add groups of cyclopropyl ether to the nuclei of other molecules include generally known chemistry and are well within the scope of the invention. gum of the technique. To add the cyclopropylacet 1 group to aromatic subtypes, palladium catalyzed coupling is easily effected. A displacement reaction is carried out to add the cyclopropyl group to alkyl substituents. The compounds of the present invention may have asymmetric center tenors and may be present as sprats, racemic mixtures or as individual diastereomers, unless specifically indicated; all isome rich forms are included in the present invention. The term (•• / -) is used to encompass * optical isomers (+) or optical isomers (-) or mixtures thereof. When any variable (eg, R, aprotic solvent) occurs more than once in any step, its definition of each occurrence is independent of its definition in all other occurrences. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. As used herein, except where indicated, "alkyl" includes aliphatic hydrocarbon groups saturated by both straight and branched chain having the specified number of carbon atoms. "Halogen" or "halo" As used herein, it means fluorine, chlorine, bromine and iodine. In this invention, the cyclopropylacetylene is prepared by means of the following scheme.

E BURN I NH4CI saturated • H The overall performance is above "He 55%. Fn • R contrast, a previous method using corrosive reagents gave approximately 42% yield, and is set as follows: SCHEME I I ^ O PCI5 < ^ CI t-BuOK / DMSO ^ (42%) as was also described in L.E »Hudson et al., 3. Arn.

Chern. Soc., 94, 1158 (1972) and W. Schoberth et al., Synthesis, 703 (1972) ,. Compound A can be synthesized by the following method.

SCHEME III p-methoxybenzyl chloride sieve 4 ü / toluno TIL SCHEME (CONTINUED) Compound A Compound A is useful in the preparation and execution of selection tests for antiviral compounds. For example, compound A is useful for isolating enzyme mutants, which are excellent screening tools for powerful antiviral compounds. In addition, compound A is useful in the establishment or determination of the binding site of other antivirals to the reverse transcription of HIV, v. Gr *,., By means of competitive inhibition. In this way, compound A is a "r * o" commercial common use for these purposes. Compound A is useful in the inhibition of reverse transferase of HIV, the prevention or treatment of infection by the virus, a human de fi ciency (HIV) and the treatment of subsequent pathological conditions such as AIDS. The treatment of AIDS or prevention or treatment of HIV infection are defined as including, but not limited to, the treatment of a wide range of HIV infection states: AIDS, ARC (AIDS-related complex), both symptomatic and asmtomatico, and real or potential exposure to VTH. For example, the compound of this invention is useful in the treatment of HIV infection after suspected previous exposure to HIV by, eg, blood transfusion, exchange of body fluids, bites, accidental needlestick, or exposure to the blood of a patient last surgery. The particular advantage of Compound A is its potent inhibition against HIV reverse transcriptase which has been rendered resistant to other antivirals, such as L-697,661, which is 3 - (C (4,7-di-chloro-1, 3-benzoxazole-2 -1 1 Irnetyl 1 -ai not) -5 -e <1-h-met Ll-p? Pd? N-1 (lH) -one; or L-695,229, which is 3-C2 - (1, 3-benz: oxaz '.? l-2 ~ 11) ethyl] -5-e +? l-6-met? lp? r? dm-2 (1H) -one, or A7T. Compound A can be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intresternal injection or infusion techniques), by aerosol inhalation, or rectally, in unit dose formulations-containing vehicles, adjuvants and conventional non-toxic pharmaceutically acceptable excipients In this way, in accordance with the present invention, a treatment method and a pharmaceutical composition for treating HIV and STDA infection are also provided. It is possible to administer to a patient in need of such treatment a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of the present invention. These pharmaceutical compositions can be * in the form of suspensions or tablets orally administrable; nasal sprays; sterile injectable preparations, for example, as sterile injectable aqueous or oleaginous suspensions, or suppositories. When administered orally as a suspension, these compositions are prepared according to techniques well known in the pharmaceutical formulating art and may contain micro-glass cellulose to impart volume, sodium acid or sodium alginate as suspending agent, rnet L-cellulose as a viscosity enhancer, and edulcorants / flavor-izant agents are known in the art. As immediate release tablets, these compositions may contain microcpstalin cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and / or other excipients, binders, extenders, disintegrants, diluents and lubricants are known in the art. When administered by aerosol or nasal inhalation, these compositions are prepared according to techniques well known in the art of the pharmaceutical formulation and which are prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption promoters for increase the biologic biodisp, fluorocarbons, and / or other solubilizing or dispersing agents known in the art. Injectable solutions or suspensions may be formulated according to the known art, using suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1,3-butanedione, water, Ringer's solution or isotonic sodium chloride solution. , or suitable dispersing or wetting agents, such as soft, fixed, sterile oils, include synthetic mono- or diglycerides, and fatty acids, including oleic acid. When administered rectally in the form of suppositories, these compositions can be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glycerol esters or polyethylene glycols, which are solid at ordinary temperatures, but are liquefy and / or dissolve in the recl cavity to release the drug. Compound A can be administered orally to humans on a dosing scale of 1 to 100 mg / kg body weight in divided doses. A preferred dosage scale is 0.1 to 10 rng / kg body weight, orally in divided doses. Another preferred dosage scale is 0.1 to 20 mg / kg body weight orally in divided doses. For combination therapy with nucleoside analogs, a preferred dosage scale for the compounds of this invention is 0.1 to 20 g / kg body weight, administered orally in divided doses, and 50 mg to 5 g / rn by weight. body of the nucleoside analogs, administered orally in divided doses. However, it is understood that the specific dose level and frequency of dosing of any particular patient may vary and that it depends on a variety of factors including the activity of the specific compound used, the metabolic stability and the duration of action. compound, age, body weight, general health, sex, diet, mode and schedule of administration, rate of excretion, combination of drugs, severity of the condition in particular, and therapy to which the guest is somito.

EXAMPLE I Preparation of cyclopropylene lacet MH4CI saturated - = • H C5H6 MW = 66 Materials Cant mmoles P ".

-Chloro-1 -pent 1 not 10 g 98 102.57 n-BuLi / cyclohexane (2.0M) 112 rnl 244 Cyclohexane 80 ml NI-C1 saturated 0 rnl To a solution of 5-chloro-1-ene in cyclohexane (80 ml) at 0 ° C under 2, n-but? Ii was added? io in cyclohexane (2.0 M, 122 ml). The mixture was heated at 75 ° C for 5 hours.

The addition of n-butyl lithium to the cinchona was exothermic; the temperature was kept below * 5 ° C during this < -additions using a H2-H2 bath. The progress of the cyclization step was monitored by means of HPLC. The reaction was considered complete when the test performance was > 90% PHLC conditions: column phenyl, CH3CN: agu: phosphoric acid, 50: 50: 0.1 eluci n 1 dur-ante socr tica 20 minutes, flow = 1.0 ml / min, UV detection at 195 n, starting material t_R =? 5 in, ci clopropiiaceti log tR-f, .p min. The product has a response factor that was 20 times greater than the starting material. Once the cyclization step was completed, the reaction was cooled to 0 ° C and quenched with saturated NH 4 Cl. The test of the organic phase with HPt C showed 5.5 < of cyclopropylacety or (85% yield). The product was purified by means of fractional distillation through a column of 15.2 c X 1.3 crn, packed with glass beads of 4 nm. The fraction with a boiling point between 45 and 75 ° C was collected. This yielded 4.2 g (65%) of cyclopropylacet 1 log as a colorless oil.

EXAMPLE 2 Preparation of 4-chlorophenyl-pivalamide Mat rials Cant "MMoles P..M. 4-Clor-oan? L? Na 75 g 596 127.57 Pivaloyl chloride 74 nl 600 120.58 (d- = 0.979) Tol ueno 600 mi NÜ2CO3 saturated 95 r l Water «1.1. 225 inl To a solution of 4-chloroan-1-ene (76 g) in toluene (600 mL) was added a2C3 (95 mL) saturated. The batch cooled to ° C and pivaloyl chloride (74 ml) was added dropwise to 45 minutes. The batch was stirred at 5-10 ° C for 60 minutes while the progress of the reaction was monitored by me «1? or HPI C. The addition of pivaloyl chloride to the aniline was exotepni ca.

HPLC conditions: column 0-8, CH3 CN; gua: phosphoric acid; Elution gradient from 40: 60: 0.1 to 80: 20: 0.1 for 20 minutes, flow - 1.0 ml / rnin, UV detection at 245 nm, starting material tR = 7.2 rnin, pivalamide tR - 12.6 inin. The product was isolated by means of filtration and washed with water «J.i. (3 x 75 mL) and dried in air with suction for 10 minutes. The product was dried in a vacuum oven at 40"C with a purge of N for 16 hours to produce 108.5 g of the product as fine white needles (86%).

EXAMPLE 3 Preparation of 4-chloro-ketoaniline C8H5F3CINO P.M. = 223.58 Materials CCaanntt .. mmmmooll oess P.M 4 - . 4 -Clo ro feni 1 pival m i da 1 100 gg 4 477..22 211.69 n- BuLi / hexane (2.5 M) 38 mi 95 Tri 1 ethyl acetate (-1.194) 6.7 rnl 56, .6 142.08 THF 75 mi Ethanol 90 rnl HCl 6N 50 mi 240 Hexane 90 mi NaOH 2M 15 mi Water d. i. 350 rnl In a 3-neck colony of 500 ml the pi valamide (10 g) was dissolved in THF (75 ml) and the mixture was cooled to 0 ° C. To this solution, n-BuL i / hexane (2.5M, 3B rnl) was added dropwise, while the internal temperature was increased to + 15 ° C. The batch was allowed to stand at 0 ° C for 2 hours. The addition of the first-equivalent of n-BuLi < •• «Pivalamide was highly exothermic. The exoteric fl was controlled by the rate of addition. To the resulting pale yellow suspension was added pure ethyl trifluoroacetate (6.7 ml), while allowing the internal temperature to rise to 10 ° C. The progress of the reaction was monitored by means of HPLC. HPLC conditions: column C-3, CH3 CN: gua: phosphoric acid; Elution gradient from 40: 60: 0.1 to 80: 20: 0.1 00 for 20 minutes, flow - 1.0 nl / rnin, UV detection at 245 nm, starting pivalarnide R =. 12.6 rnin, cetopí valarm da R _ 11. b? n? n.6. There was typically 85A% and 10-15A% of unreacted pivalarnide. The reaction was quenched by adding 6N HCl (10 rnl) and water d. i. (20 mi). The HPLC test at this point showed 13.1 g (90%) of product. The sol was concentrated in vacuo to approximately 50 mL and ethanol (50 mL) to remove hexane and THF. To the batch was added 6N HCl (40 mL) and the mixture was heated to reflux (80 ° C) for 1 hour. The HPLC test showed 85-90A% of the ketoaniline, and 10A% of unreacted pivalaride. In this way, the acylated material undergoes hydrolysis while the unreacted pivalarnide remains unchanged. The test performance at this point was 7.78 g (74%). The batch was concentrated in vacuum until approximately 50 ml, when a precipitate was formed (presumably the HCl salt of the product). Distillation was stopped and the batch cooled to 0 ° C. After 1 hour of rest, the batch was filtered and washed with hexane (3 x 30 rnl). The hexane washes removed the unreacted pivalanide product. The solid was analyzed by HPLC to ensure that it had been completely removed at this point. The filtrate and the washes typically contain 1.2-1.5 g of the product (8-12%). The majority of the product loss was in the aqueous filtrate. Dry salt in a vacuum oven at 40 ° C for 16 hours to produce 10.4 g of a solid that was 71.4% pure by weight (70% yield). The salt was suspended in water d.i. (260 ml) and neutralized to a pH of about 6-7 with 2M NaOH (15 m). It was critical not to bring the pH above 9.0 due to the decomposition of the product. The resulting bright yellow solid was isolated by filtration and washed with water d.i. (2 x 25 rnl). The product was dried in a vacuum oven at 40 ° C for 16 hours to yield 6 g of cefoaniline which was 96.6% pure by weight (54% yield). The product was purified adi citonally by rectanzation of hexane.

EXAMPLE 4 Preparation of N-4-methoxybenzyl-ketoanine C1SH13F3CIN02 P.M. = 343.735 Materi les Cant inmole1 P.M, Ce anili ta 15.5 «j 69.5 223.58 Chloride «le p-rnet oxi bencí 1 or 10.9 g 69, .8 Molecular sieves« Je 48 90 g Toluene 70 mi Ace ona 500 rnl He no 120 rnl In a 250 ml flask, the ketoaniline (15.5 g) was charged, fi-activated molecular sieves (50 g) and toluene (75 ml). The mixture was stirred at 23 ° C under N for 24 hours. The test by * HPI. C showed a mixture of approximately 1: 1"Je product and starting material. HPLC conditions: C-8 column, CH3CN: gua: phosphoric acid, 1 Socratic elution at 65: 35: 0.1 for 20 minutes, flow = 1.0 l / inin, UV detection at 260 n, toluene -5.7 min, ketoaniline starting time tR = 6.5 min, product tR = L5.0 inm. There was typically 25A% toluene. The reaction was loaded with fresh molecular sieves (40 g) and stirred for an additional 3 days at 23 ° C. The reaction was considered complete when "I am less than 2A% of the starting material. The mixture was filtered through celite and washed with acetone (7 x 75 rnl) until most of the yellow color was washed out of the celite. The filtrate was concentrated to yield 27 g of an orange yellow oil which solidified upon standing. The solid was purified by dissolving it in hot hexane (100 ml). The batch was cooled to room temperature, then to 0 ° C in a H 2 -el bath. After a rest of 1.5 hours, the batch was filtered and washed with cold hexane (2 x 10 ml). The batch was air-dried with 10-minute hard-suction, then dried in a vacuum oven at 40 ° C for 2 hours. This produced 20.5 g (86%) of a bright yellow powder.

EXAMPLE 5 Preparation of the Anoalcohol C21H19F3CIN02 P.M. = 409 Materials Cant mmoles P. ..

Ce ona 175 g 0.51 343 IR, 2S-N- ?? rrol i dini 1 norefedpna 264 ing 1.3 205 Ciclopropí laceti leño (d-0.8) 0.1 rnl 1.21 66 sec-BuLi (1.3 M in cyclohexane) 2 mi 2.6 THF (KF = 20 ug / ml) 4 mi Citric acid 1M 10 rnl Acetic acid ethyl acetate 6 ml The ?? rroli dimlefedri na (264 rng) was dissolved in THF (2 mL) and the mixture was cooled to -5 ° C. To the mixture was added, drop by drop, at -5 ° C and under 2, pure cyclopropylacet (0.11 rnl) and sec-but illi t 10 (2.0 ml). The mixture was allowed to stand at -5 ° C for 30 minutes and then cooled to ~ 45 ° C. The addition of sec-butyl itao caused an exotepna that was maintained between -5 and 0 ° C, regulating the rate of addition. The ketone (175 mg) was dissolved in THF (1.0 ml) under 2 and added to the ammonium mixture for 2-3 minutes allowing the internal temperature to rise to ~ 40 <.;: "C during the addition, the resulting solution light orange color or stand at -40 ° C for 60 minutes and was quenched by adding 1M citric acid (3 ml) and ethyl acetate (3 ml). The reaction was warmed to room temperature and the layers were separated The lower layer of water was extracted with ethyl acetate (3 mL), the combined organic layers were washed with 1M citric acid (2 x 3 minutes). it was tested by means of HPLC to determine the conversion percentage and EE of the product HPLC conditions: column C-8, CH3 CN: gua: phosphoric acid, 1: 65 elution 65: 35: 0.1 for 20 minutes, flow = 1.0 ml / rnin, detection at UV at 245 nm, material «Je departure tR = 12.8 min, product R = 10.3 rnin Chiral HPLC conditions: column of stationary phase of araylose, hexane: isopropanol 85:15, eluci nsocratica, flow- 1.0 nl / rnin, UV detection at 252 nrn, material pr = 4.9 rnin, enant lornero greater tR - 5.5 rnin, enant 1 omero minor t_R = 25.0 inin. The enant lornép co excess was 98% and the conversion of the reaction was 93% (6A% of the starting material). The performance of the test was 92%.

EXAMPLE 6 Mate i les Cant. Immoles P..M.

Arni no alcohol 3.2 g 7.8 409 Phosgene in toluene (1.93M) 4.6 mi 8.89 Trieti lamina (d = 0.726) 5.4 rnl 39 101 THF (KF <100 μg / rnl) .1.5 rnl Water d.i. 15 rnl. EtO c 45 ml Hexane 30 ml citric acid 1 M 40 rnl Saturated brine 25 ml F 1 arninoa 1 cohol was dissolved in THF "(15? Nl) and cooled to -10 ° C under N2. To the mixture was added tpeti lami a (5.4 rnl) and phosgene in toluene (4.6 i) The addition of phosgene caused an exotherm to be maintained below 20 ° C regulating the rate of addition.The progress of the reaction was monitored by HPLC and was typically Complete at 15 minutes HPLC conditions: column C-8, CH3CN: gua: phosphoric acid, elution gradient 50: 50: 0.1 to 90:10:01 for 20 minutes, flow = 1.5 ml / inin, detection in UV at 252 nrn, starting material tR = 14.6 min, product tR = l6.0 rnin The reaction was cooled to 0 ° C and quenched with ice-cold water (15 ml) and ethyl acetate (20 ml). Saturated brine was used to break up any emulsion, the organic layer was removed and the aqueous layer was extracted with ethyl acetate (15 mL). gone citric IM (40 ml) and saturated brine (25 ml). The organic layer was dried (N 2 0") and concentrated in vacuo to yield 3.8 g of a brown oil. The product was crystallized from hexane: ethyl acetate 5: 1 (25 nmol), cooled to 0 ° C, left to stand for one hour and filtered. The cake was washed with hexane: ethyl acetate : 1 cold (2 x 5 rnl). The cake is air dried with suction to yield 2.9 g (85%) of a light orange solid.

EXAMPLE 7 Preparation of Compound A Compound A Materials Quant. Mmoles P.M.

Compound A PMB, 7 0.8 g 1.83 435 Citrus ammonium nitrate 4.'- g 8.0 548.23 CH3CN 15 mi Acetate of ethyl 30 1 Water d.i. 30 rn.1 Saturated brine 1 i Compound A p-rnetoxybenzyl was dissolved in OH 3 CN (15 ml). To this solution was added a solution of ceric ammonium nitrate (4.4 g) in water (5 mL). The reaction was typically complete in 2 hours at 23 ° C, as determined by HPLC. HPLC conditions: column 0-8, CH3CN: water: phosphorus acid, gradient of 50: 50: 0.1 to 90:10:01 for 20 minutes, flow - 1.5 l / min, «UV detection to 252 nm, torque material tR- 16.0 mm, product tR = 9.0 m. The reaction was diluted with water d.i. (5 ml) and concentrated to approximately half the volume. The product was extracted from the resulting aqueous layer with ethyl acetate (2 x 15). The combined organic extracts were washed with water d.i. (2 x 10 rnl) and brine (10 rril). The organic layer was concentrated under vacuum to produce a yellow gum. The product was isolated by chromatography on silica gel.

EXAMPLE 8 N- (4-chlorophenyl) -2,2-dimethylpropanamide To a 3-necked round bottom flask of 51 with a stirrer on top, 4-chloroan was added. lina (127.57 g, 1 mol), 1200 ml of CHCl3, V 1200 nl of saturated aqueous solution of Na C 3. An addition funnel was connected to the flask and charged with 2, 2-dirnethylpropanoyl chloride. mi, 1.05 moles). The acid chloride was added dropwise to the stirred mixture visually for 1 hour. The resulting mixture was stirred at room temperature for 23 hours and added thereto. Part of the product was separated from the mixture as white crystals. These crystals were collected by means of filtration. The filtrate was transferred to a separatory funnel and the layers were separated. The chloroform layer was washed with water and brine. Drying (MgSO ^), filtration and elimination of the solvent in vacuo gave additional product. The portions of the product were combined and recrystallized from boiling EtOAc-hexane to give 185.6 g of N- (4-chlorophenol) -2,2-d-methylpropanamide as a solid. i no b 1 n co.

J EXAMPLE 9 (-) 6-Chloro-4-cyclopropylethynyl-4-trifluoromethyl-1, -dihydro-2H-3, 1-benzoxazin-2-one (Compound A) and (+) 6-Chloro-4-cyclopropyl-ethynyl -4-trifluoromethyl-l, 4-dihydro-2H-3, 1-benzoxazin-2-one Step A 2 (2 -Arní no -5-clor-of eni 1) -4-c? clopropyl -1,1,1-tpf luoro-3-but i n-2 -ol A solution of cyclopropylacetide of bro or agnesium was prepared from 23 g of cyclopropylacetylene (0.38 mole) in 250 m of THF per ml. of the dropwise addition of 116 ml of a 3.0M solution of ethyl agnesium bromide in ether (0.348 moles) for 1 hour. This solution was maintained at 0 ° C for 1 hour, then at 40 ° C for 3 hours. To this solution, re-heated at 0 ° C, 15.56 g of l- (2-a? O -5-chlorophenyl) -2, 2, 2 -tri f luoromet i letanone (0.0696 moles), as a solid, for 5 minutes. The reaction mixture was allowed to stir at 0 ° C for 1.5 hours. The reaction was quenched at 0 ° C by the dropwise addition of 700 ml of saturated aqueous solution of ammonium chloride. The mixture was extracted with 2 x 400 rnl portions of ethyl acetate, the combined organic phases were washed with brine and dried over g. The removal of the drying agent and the solvents left a yellow solid »This material was recrystallized from boiling hexane (100 rnl final volume) to yield 14.67 g of 2- (2-arn? No ~ 5-chlorofem 1) -4-c? Cloprop? Ll, 1, l-tp f luoro-3-but? N-2 - ol. A second harvest (2.1 g) was obtained for the concentration of mother liquors. P.f. : 153-154 ° C 1 H-NMR (CDCl 3): & 0.84 (rn, 2H, 0.90 (rn, 2H (, 1.38 (m, 1H), 4.50 (br s, 3H, 6.69 (d, 3 = 8.5 Hz, 1H), 7.13 (dd, 3 - 2.5, 8.5 Hz, 1H), 7.55 (d, J = 2.5 H, IH). P o S O () 6-C1-oro-4-cyclopropylethi nor 1 -4- tp fl uoronet 11-1,6-dihydro-2H-3, 1-benzoxa 1 n-2 -one A solution of 2- (2-am? No-5-chlorophen? 1) -k -c? Clopro? L-1,1,1-tpfluoro-3-but? N-2-ol (15.00 g, 0.0518 mol and 41.98 g ( 0.259 moles) of 1, l '-carbonyl di-jolzole in 250 ml of dry FHF was stirred under argon at 55 ° C for 24 hours.The solvent was removed on a rotary evaporator and the residue was divided between 500 rn. ethyl and 400 ml of water.

The layers were separated and the aqueous phase was extracted once more with ethyl acetate. The combined ethyl acetate extracts were washed with 2 x 200 mL of 2% aqueous HCl, saturated NaHCO 3, aqueous, and brine. Drying over MgSO, filtration and removal of the solvent in vacuo gave 16.42 g of the title compound as a solid. Recrystallization from ethylhexane acetate yielded 12.97 g of (±) 6-chloro-4-c-chloropethyl-4-trifluoride-1, 4-d? H? analytically pure dro-2H-3, 1- benzoxaz m-2-one, as white crystals. P.f .: 178-180 ° C. * H NMR (CDCI3): 0.85 (m, 2H), 0.94 (rn, 2H), 1.40 (rn, IH), 5.81 (d, J ^ 8.5 Hz, 1H) 7.37 (dd, 3 = 2.5, 8.5 Hz , 1H), 7.49 di * ("1.3 = 2.5 Hz, 1H), 8.87 íbr s, 1H). Step C 6 - Chloro- 1 - (1S) -alphafo and 1-4-cyclopropyl my 1-4-tp fl uororneti 1-1,4 -dihi dro-2H-3, l-benzoxazn-2-one A a solution containing (±) 6-chloro-4-c? clo? rop? l-et 1 m 1-4-tr? fluorornet? ll, 4- < iih? dro-2H-3, l-benzoxaz? n-2-one (12.97 g, 0.041 mol), 4-dimethylarninopy pdina (1.02 g, 0.0083 mol) and aculo (-) camphor chloride 1 ( 14.22 g, 0.06556 moles) in 350 ml of dry dichloromethane, stirred under argon in an ice bath, triet 1 sheet (22.84 rnl, 0.164 moles) was added. The cooling bath was removed and the reaction was allowed to proceed at room temperature. "After 75 minutes, the reaction was considered complete by means of thin layer chromatography (S1O2, 4% EtOAc in CHCl3), and the solution was diluted with 500 ml of CHCl3, then washed with acid. 10% citric acid (2 X), water (IX), and brine (IX) .Drying (MgSO /;), filtration and elimination of the solvent under vacuum left a colorless foam.This material was crushed with 200 rnl After cooling to room temperature, the desired diastereomeric camphorate bound was precipitated.The solute was collected on a glass material, washed with a little cold hexane and dried in vacuo to give 7.70 g of 6-fold. chloro-l ~ (1S) -alphafor? l-4-c? clo-? ro ?? le 1 or 1-4-tpfluorometi ll, 4 ~ d? h? dro-2H-3, l- benzoxaz i n- 2 -one as white crystals, Pf: 164-1650 C. Purity by HPLC: 99.2% @ 254 nm. 1 H-NMR (CDCl 3): or 0.77 (s, 3H), 0.86-0.96 (rn, 4H), 1.08! 6 (s, 3H), 1.19 (s, 3H), 1.44 (rn, 1H), 1.76 (rn, 1H), 1.95 (in, 1H), 2.51 (m, 2H), 7.42 (dd, 3 = 2.4, 9.0 Hz, 1H), 7.63 (rn, 2H). Step T (-) 6-Chloro-4-c? clopropí let i n? l-4 - t rif 1 uoroinot i l, -dihidr-o- 2H-3, 1-benzoxaz? n-2-one (Cornpuest or A) 6-Chloro-1- (1S) -alpha was dissolved for i 1 -4-cyclopropyl let? n-4-tpfluorom t? 1-1, -d? hi ro -4 (H) -, 1 -t) en-? oxa- ~ 'i n-2-one (7.50 g, 0.01512 mol) in 150 ml of n-butanol < * 60 ° C ba or an argon atmosphere. To this solution were added ml of 1N HCl. This solution was maintained at 60 ° C for 72 hours. The mixture was neutralized with aqueous NaHC 3 and the n-butanol was removed in vacuo. The residue was dissolved in 150 ml of THF and treated with 50 ml of 2N LiOH for 3 hours at room temperature. This mixture was diluted with ethyl acetate and washed with the portions of water and one of brine. Drying (MgSO), filtration and elimination of the solvent in vacuo gave a white solid. This material was recrystallized from hot hexane to give 3.43 g of 1 -) 6-chloro-4-c-clopropyl-1-yr-1-trifluoromethane, 4-dihydro-2H-3, 1 - benzoxazm-2-one, as white crystals. MP: 131-132 ° C; [»D20 = -84.7 ° (CHC13, c-p.005 g rnl-l); 1 H-NMR (CDCl 3): d 0-85 (rn, 2H), 0.94 (rn, 2H), 1.40 (, 1H), 6.81 (d, 3 = 8.5 H, 1H), 7.37 (dd, 3 = 2.5, 3.5 Hz, 1H), 7.49 (d, 3 = 2.5 Hz, 1H), 8.87 (br, s, IH).

P so F. (») 6 -Cioro-4-c? Clopr-op? let i ni 1 - 4 -tpf luoromet? l - 1, 4-di huJro-2H-3, 1-benzoxaz i n-2-one The mother liquors from step C above were purified by means of column chromatography on silica gel using 10% ethyl acetate in hexane as eluent. The undesired pure diastereomer (a colorless foam) was hydrolysed according to step D. The benzoxazinone enant lornérica, (*) 6-chloro-4- ci clopropyl let was obtained. nil -4- t ri f 1 uorornet 11 - l, 4-d? h? dro-2H 3, 1-benzoxazin-2-one, as white crystals. P. f: 131-132 ° C; Ca3D20 _ = »84.4 ° (CHCl3, c = 0.005 g rnl-'H; 1H-NMR (CDCI3): d 0.85 (rn, 2H), 0.94 (m, 2H), 1.40 (rn, IH), 6.81 (d , 3 = 8.5 H, 1H), 7.37 (dd, 3 = 2.5, 8.5 Hz, 1H), 7.49 (d, 3 = 2.5 H, 1H), 8.87 (hr, s, 1H).

Reverse Pseudo Transease Test The test measures the incorporation of deoxidized guanosme rnonophosphate by means of HIV recirculating reverse transacpttase (TIR VTH) (or other IT) into acid-precipitable cDNA at the dGTP and poly r K values ( C) »oligo d (G) 12-18. The inhibitors of the present invention inhibit this 1 ncorpo acid. The tests were carried out in tris 55 rnM (H 8.2) -KCl 30 mM-MgCl2 30 rnM-dithiot reaol lmM ~ 20ug of rC: dGi2-i8 (Pharmacia) by? Nl-8 rnM [3H] dGTP (New England Nuclear) -0. 1% Triton X- 100-50 M etiiengl icol-bis (ß-aimno-et 1 lico ether) - N, N, N ', N' - tetraacetic acid (FGTA) -l mg of bovine serum albumin by * rnl. After 60 minutes' incubation at 37 ° C, acid-precipitable material was collected on glass fiber filters using a semiaut or attic cellular harvester. The bacterial cell extracts containing IT were diluted to within a linear scale of the test, and activity was determined in the presence and absence of inhibitor. The heterodimer of purified HIV-1 IT produced in E. coli also served co or control. The results were determined as concentration of inhibitor to give 50% inhibition (IC50 wt), in nano? Noles / 111 ro. Compound A gave a TC50 t of 2 nM. For the double-negative test (drn), IT was used to the 7 in the test. TI A17 is resistant to different nopyridones, as described in Nunberg, 3.H. and others. , 3.

V 11 -o 1., 65, 4887 (1991). The results were measured as IC50 drn on nannols / 1 nit. Compound A gave a wt IC50 of 85 nM.

Cell Extension Test Inhibition of VTH extension in cell culture was carried out according to Nunberg, 3.H. and others. , 3. V 1 ro1. , 65, 4887 (1991). In this test, MT-4 lymphoid cells were infected with HIV-1 (wild type, at least as indicated) using a predetermined inoculum, and the cultures were incubated for 24 hours. At this time, <1% of the cells were positive by means of indirect immunofluorescence. Afterwards, the cells were washed extensively and distributed in 96-well culture dishes. Duplicate serial dilutions were added to the wells, and the cultures were continued for 3 additional minutes. At 4 days after infection, 100% of the cells were infected in the control cultures. The p24 accumulation of HIV-1 was directly correlated with the spread of the virus. The inhibitory concentration of cell culture was defined as the inhibitor concentration in nanorols / 111 I? that reduced the spread of the infection by at least 95%, or Although the foregoing specification would teach the principles of the present invention, with examples provided for the purpose of illustration, it will be understood that the practice of the invention encompasses all variations, adaptations or customary uses "fall within" the scope of the following claims. and its equivalents.

Claims (7)

NOVELTY OF THE INVENTION CLAIMS

1. - A procedure for the preparation of cyclopropylacetylene, < It was purchased in the steps of (a) mixing at least about 1.0 equivalents of base strength chosen from the group consisting of n-but 11111, sodium amide, diethyl sodium, hydride. sodium, potassium hydride, b? s (tpmetilsi 11 sodium daeda, b? s (tprnet ilsil 11) arn? < potassium hydride, dusopropylarn lithium, sec-but lili uncle, tert-butyllithium, and tet The concentration of lithium in lithium in an aqueous solvent with an equivalent of 5-halo-1-pentene in aprotic solvent at a temperature between about -20 and about 150 ° C (b) the temperature of the reaction mixture to a scale of between approximately 0 to approximately 150 ° C and maintaining the temperature within the scale for at least about 15 minutes, or until the cycle is substantially complete; ) extinguish the reaction with a source of protons,

2. a preparation process for cyclopr-opyl acetic acid, which comprises steps of (a) nezclai of at least about 1.0 or «| u? valent of strong base chosen from the group < } Which consists of n-but i 1 iit 10, sodium amide, sodium diethylamide, sodium hydride, potassium hydride, bi s (t runetilsi lil) am? Sodium, b? s (methyl silyl) potassium oxide, lithium diisopropyl ida, sec-butylite 10, tert-butyl 11? t 10, and lithium tet ramethi piperidide in an appropriate solvent with an equivalent of 5 ~ halo-1-penne in aprotic solvent at a temperature of about -20 and about 150 ° C; (b) allowing the temperature of the reaction mixture to rise to a scale between about 0 and about 150 ° C and maintaining the temperature within the scale for at least about 15 minutes, or until the cyclization is substantially complete; (c) cooling the reaction mixture to a temperature between about -30 and about 50 ° C; (d) quenching the reaction with any proton source.

3. The method according to claim 2, further characterized in that it comprises purifying the desired product of the clopiroleum and wood.

4. The method according to claim 1, further characterized because the strong base is n-bu? Ll itio.

5. The process according to any of claims 1-3, characterizes «Jo to« jernas because the aprotic solvent is selected «je THF, 2, 5-d? Met? L -THF, 1,4- dioxane, MRBE, di ethoxy methane, di ethoxyethane, cyclohexane, hexane, and hexane with tetramethylenediamine.

6. The process according to claim 5, further characterized in that the aprotic solvent is cyclohexane.

7. - The process according to any of claims 1-3, further characterized because the source of protons is selected from NH "C1 saturated, HCl and H2 O *. H. The process according to any of claims 1-3, further characterized in that the 5-halo-1-pentyne is 5-chloro-1-pentane. 9.- A procedure of preparation of ci < , I made the laceti which comprises the steps of (a) mixing between about 2.0 and approximately 2.5 equivalents of n-butyllithium in cyclohexane with an equivalent of 5-chloro-1-pentmo in cyclohexane at about 0 ° C; (b) heating the reaction to about 75 ° C and maintaining the reaction at that temperature for about 5 hours, or until the cyclization is substantially complete; (c) cooling the reaction mixture to about 0 ° C; (d) quenching the reaction with saturated NH.sub.1; 10. The process according to claim 9, further characterized in that it comprises purifying the desired product of cyclopropylacet 1 logs.