WO2022037469A1 - Long-acting relaxin-2 analog - Google Patents

Abstract

Provided is a long-acting relaxin-2 analog, relating to the field of pharmaceutical synthesis. The analog is used for preparing a pharmaceutical composition for treating diseases. The pharmaceutical composition is used for treating various diseases, and the various diseases comprise various heart failure and various inflammatory diseases.

Description

A long-acting relaxin 2 analog

This application claims the priority of the Chinese patent application with the application number of 202010826963.2 and the invention titled "A kind of long-acting relaxin 2 analog" filed with the China Patent Office on August 17, 2020, the entire contents of which are incorporated herein by reference Applying.

technical field

The invention relates to the field of pharmaceutical synthesis, in particular to a long-acting relaxin 2 analog and use thereof.

Background technique

Relaxin 2 was first discovered by Frederick et al. in 1926 when they studied changes in the pelvic girdle during pregnancy. It is a natural peptide hormone, named after the significant relaxation of the pubic ligament in pregnant women. Blood vessels are the target organs of relaxin.

The heart can produce relaxin 2 and exert cardioprotection and extracellular matrix regulation through local receptors. There is relaxin 2 receptor mRNA expression in human atrium/ventricular, and its density is higher than that of myometrium. Atrial and ventricular myocardium in patients with heart failure The expression of relaxin 2 is persistently high, and it is related to the severity of heart failure. The role of relaxin 2 in the treatment of acute heart failure is to dilate systemic blood vessels, dilate renal blood vessels, and increase arterial compliance.

Relaxin 2 is involved in hemodynamic regulation, maintains circulatory homeostasis, induces NO production, reduces vascular tone, antagonizes vascular contraction responses to ET-1, Ang II and catecholamines, and increases cardiac and renal blood flow; affects water intake and The regulation of renal function maintains volume balance; participates in the remodeling of the vasculature, improves arterial elasticity; stimulates the synthesis and release of atrial peptides, inhibits fibroblast activation and proliferation, and inhibits myocardial hypertrophy; resists myocardial ischemia/reperfusion injury and reduces heart rhythm Abnormal; involved in repair and myocardial regeneration after myocardial infarction; promote collagen degradation, prevent or reverse cardiac fibrosis.

Clinically, relaxin 2 is mainly used for the treatment of patients with heart failure, and is also used for various inflammatory diseases. However, due to the short half-life of relaxin 2, patients need to be administered every day, and once a week cannot be achieved. The purpose of the present invention is to provide a long-acting relaxin 2 analog with longer half-life.

SUMMARY OF THE INVENTION

In view of this, the present invention provides a long-acting relaxin 2 analog and use thereof.

In order to achieve the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

The present invention first provides a compound whose structure is shown in formula I, a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex formed by the compound, and a prodrug based on the compound, or any mixture of the above forms.

When X1 in formula I is S, X2 is S, or is CH2;

When X1 in formula I is CH2, X2 is S, or is CH2;

X1 in formula I is NH, and X2 is CO;

When X1 in formula I is CO, X2 is NH;

AA1 in formula I is any codable amino acid except Cys and Glu, or any non-codable amino acid without SH group;

AA2 in formula I is Asp, or Asn, or Glu, or GIn, or GIn, or Ser, or Thr;

AA3 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

AA4 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

AA5 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

AA6 in formula I is any codable amino acid except Cys, or any non-codable amino acid without SH group, or is absent;

AA7 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

AA8 in formula I is any codable amino acid except Cys, or any non-codable amino acid without SH group, or is absent;

AA9 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

AA10 in formula I is Lys, or Dah, or Orn, or Dab, or Dap;

AA11 in formula I is NH2, or is OH;

R in formula I is cholesterol succinate monoester, or 2-cholesteryl acetic acid, or 2-cholesteryl propionic acid, or 3-cholesteryl propionic acid, or 2-cholesteryl butyric acid, or 2-cholesteryl isobutyrate acid, or 3-cholesterylbutyric acid, or 3-cholesterylisobutyric acid, 4-cholesterylbutyric acid, or 2-cholesterylvaleric acid, or 2-cholesterylisovaleric acid, or 3-cholesterylvaleric acid, Either 5-cholesteryl valeric acid, or 2-cholesteryl caproic acid, or 6-cholesteryl caproic acid, or 2-cholesteryl heptanoic acid, or 7-cholesteryl heptanoic acid, or 2-cholesteryl caproic acid, or 8 -Cholesterol octanoic acid, or HO2C(CH2)n1CO-(γGlu)n2-(PEGn3( _CH2 )n4CO)n5-;

Where: n1 is an integer from 10 to 20;

n2 is an integer from 1 to 5;

n3 is an integer from 1 to 30;

n4 is an integer from 1 to 5;

n5 is an integer from 1 to 5;

When AA11 in formula I is OH, X1 is S, and X2 is CH2, R can be absent;

When AA11 in formula I is OH, X1 is CH2, and X2 is S, R can be absent;

When AA11 in formula I is OH, X1 is CH2, and X2 is CH2, R can be absent;

When AA11 in formula I is OH, X1 is NH, and X2 is CO, R can be absent;

When AA11 in formula I is OH, X1 is CO, and X2 is NH, R can be absent;

When AA11 in formula I is NH2, X1 is S, and X2 is S, R can be absent;

When AA11 in formula I is NH2, X1 is S, and X2 is CH2, R can be absent;

When AA11 in formula I is NH2, X1 is CH2, and X2 is S, R can be absent;

When AA11 in formula I is NH2, X1 is CH2, and X2 is CH2, R can be absent;

When AA11 in structure I is NH2, X1 is NH, and X2 is CO, R may be absent;

When AA11 in formula I is NH2, X1 is CO, and X2 is NH, R may be absent.

The present invention also provides pharmaceutical compositions comprising compounds according to the present invention, as well as providing pharmaceutical compositions of compounds of the present invention for use in the manufacture of pharmaceutical compositions for the treatment of diseases.

Use of the pharmaceutical composition of the present invention for treating various diseases, including heart failure and various inflammatory conditions.

Further, the various types of heart failure include acute congestive heart failure, compensated heart failure and the like.

Further, the various inflammatory conditions mainly include eosinophilic airway hyperresponsiveness, asthma, rheumatoid arthritis, multiple sclerosis, ankylosing spondylitis, inflammatory bowel disease, gout, myositis, systemic lupus erythematosus , vasculitis, sepsis, trauma, wound healing, eczema, dermatitis, scleroderma, acne, urticaria, psoriasis and allergic reactions.

More contents involved in the present invention are described in detail below, or some of them can also be realized in the embodiments of the present invention. Unless otherwise indicated, the amounts used herein to indicate the amounts of the various components and reaction conditions can be interpreted as "approximately" or "approximately" in any case. Correspondingly, unless otherwise specified, the numerical parameters quoted in the following and in the claims are approximate parameters, and different numerical parameters may be obtained due to differences in standard errors under respective experimental conditions.

In this paper, when there is disagreement or doubt between the chemical structural formula and the chemical name of a compound, the chemical structural formula is used to exactly define the compound. The compounds described herein may contain one or more chiral centers, and/or double bonds and the like, and may also exist as stereoisomers, including double bond isomers (such as geometric isomers), optically active enantiomers or diastereomers. Accordingly, any chemical structure within the scope of the description herein, whether in part or in the whole structure containing similar structures above, includes all possible enantiomers and diastereomers of the compound, including Any single stereoisomer (eg, pure geometric isomer, pure enantiomer, or pure diastereomer) and any mixture of these isomers are included. These racemic and stereoisomeric mixtures can also be further resolved into their constituent enantiomers or stereoisomers by those skilled in the art using continuous separation techniques or chiral molecular synthesis methods body.

Compounds of formula I include, but are not limited to, optical isomers, racemates and/or other mixtures of these compounds. In the above case, single enantiomer or diastereomer, such as optical isomer, can be obtained by asymmetric synthesis method or racemate separation method. Resolution of the racemates can be accomplished by various methods, such as conventional recrystallization with a resolution-promoting agent, or by chromatography. In addition, compounds of structure such as Formula I also encompass double-bonded cis and/or trans isomers.

The compounds of the present invention include, but are not limited to, the compounds of formula I and all of their various pharmaceutically usable forms. The different pharmaceutically acceptable forms of these compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the aforementioned substances and the aforementioned forms of these compounds. any mixture.

detailed description

The invention discloses a long-acting relaxin 2 analog and its use, and those skilled in the art can learn from the content of this article and appropriately improve the process parameters to achieve. It should be particularly pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are deemed to be included in the present invention. The method and application of the present invention have been described through the preferred embodiments, and it is obvious that relevant persons can make changes or appropriate changes and combinations of the methods and applications described herein without departing from the content, spirit and scope of the present invention to achieve and Apply the technology of the present invention.

The long-acting relaxin 2 analogs provided by the present invention and the raw materials and reagents used in their uses can be purchased from the market. Below in conjunction with embodiment, the present invention is further elaborated:

The corresponding Chinese names of the English abbreviations involved in the present invention are shown in the following table:

Example 1 Preparation of Compound 1

The preparation method includes: preparing a peptide resin by a solid-phase polypeptide synthesis method, and then subjecting the peptide resin to acidolysis to obtain a crude product, and finally purifying the crude product to obtain a pure product; wherein the step of preparing the peptide resin by the solid-phase polypeptide synthesis method is to pass the solid-phase peptide synthesis method on the carrier resin. The corresponding protected amino acids or fragments in the following sequences are sequentially connected by the coupled synthesis method to prepare the peptide resin:

In the above preparation method, the dosage of the Fmoc-protected amino acid is 1.2 to 6 times of the total moles of the resin charged; preferably 2.5 to 3.5 times.

In the above preparation method, the substitution value of the carrier resin is 0.2-1.0 mmol/g resin, and the preferred substitution value is 0.3-0.5 mmol/g resin.

As a preferred solution of the present invention, the solid-phase coupling synthesis method is as follows: the protected amino acid-resin obtained in the previous step is subjected to a coupling reaction with the next protected amino acid after removing the Fmoc protecting group. The deprotection time for de-Fmoc protection is 10-60 minutes, preferably 15-25 minutes. The coupling reaction time is 60-300 minutes, preferably 100-140 minutes.

The coupling reaction needs to add a condensation reagent, and the condensation reagent is selected from DIC (N,N-diisopropylcarbodiimide), N,N-dicyclohexylcarbodiimide, benzotriazole hexafluorophosphate -1-yl-oxytripyrrolidinophosphorus, 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethylurea hexafluorophosphate , benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate or O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoro One of boronate esters; preferably N,N-diisopropylcarbodiimide. The molar amount of the condensation reagent is 1.2-6 times, preferably 2.5-3.5 times, the total moles of amino groups in the amino resin.

The coupling reaction needs to add an activating reagent, and the activating reagent is selected from 1-hydroxybenzotriazole or N-hydroxy-7-azabenzotriazole, preferably 1-hydroxybenzotriazole. The dosage of the activating agent is 1.2-6 times, preferably 2.5-3.5 times, the total moles of amino groups in the amino resin.

As a preferred solution of the present invention, the reagent for removing Fmoc protection is a PIP/DMF (piperidine/N,N-dimethylformamide) mixed solution, and the mixed solution contains piperidine in an amount of 10-30% (V ). The dosage of the de-Fmoc protecting reagent is 5-15 mL per gram of amino resin, preferably 8-12 mL per gram of amino resin.

Preferably, the peptide resin is subjected to acid hydrolysis while removing the resin and the side chain protecting group to obtain a crude product:

Further preferably, the acid hydrolyzing agent used during the acidolysis of the peptide resin is a mixed solvent of trifluoroacetic acid (TFA), 1,2-ethanedithiol (EDT) and water, and the volume ratio of the mixed solvent is: TFA It is 80-95%, EDT is 1-10%, and the balance is water.

More preferably, the volume ratio of the mixed solvent is: TFA is 89-91%, EDT is 4-6%, and the balance is water. The optimal volume ratio of the mixed solvent is: TFA is 90%, EDT is 5%, and the balance is water.

The dosage of the acid hydrolyzing agent is 4-15 mL of acid hydrolyzing agent per gram of peptide resin; preferably, 7-10 mL of acid hydrolyzing agent is required per gram of peptide resin.

The cleavage time using the acid hydrolyzing agent is 1 to 6 hours at room temperature, preferably 3 to 4 hours.

Further, the crude product was purified by high performance liquid chromatography and freeze-dried to obtain pure product.

1. Synthesis of peptide resin

Using Rink Amide BHHA resin as carrier resin, through de-Fmoc protection and coupling reaction, coupled with the protected amino acids shown in the following table in turn to prepare peptide resin. The protected amino acids corresponding to the protected amino acids used in this example are as follows:

Peptide sequence n=	protect amino acids
1	Fmoc-Lys(AEEA-AEEA-γGlu(OtBu)-octadecanedioic acid mono

	tert-butyl ester)
2	Fmoc-Ser(tBu)
3	Fmoc-Trp(Boc)
4	Fmoc-Thr(tBu)
5	Fmoc-Ser(tBu)
6	Fmoc-Met
7	Fmoc-Gly
8	Fmoc-Cys(Mtt)
9	Fmoc-Ile
10	Fmoc-Ala
11	Fmoc-Ile
12	Fmoc-Gln(Trt)
13	Fmoc-Ala
14	Fmoc-Arg(Pbf)
15	Fmoc-Val
16	Fmoc-Leu
17	Fmoc-Glu(OtBu)
18	Fmoc-Arg(Pbf)
19	Fmoc-Gly
20	Fmoc-Cys(Trt)
twenty one	Fmoc-Leu
twenty two	Fmoc-Lys(Boc)
twenty three	Fmoc-Ile
twenty four	Fmoc-Val
25	Fmoc-Glu(OtBu)
26	Fmoc-Glu(OtBu)
27	Fmoc-Met

28	Fmoc-Trp(Boc)
29	Fmoc-Ser(tBu)
30	Boc-Asp(OtBu)
Test 1	Fmoc-Cys-OtBu
Test 2	Fmoc-Phe
Test 3	Fmoc-Arg(Pbf)
Test 4	Fmoc-Ala
Test 5	Fmoc-Leu
Test 6	Fmoc-Ser(tBu)
Test 7	Fmoc-Arg(Pbf)
Test 8	Fmoc-Lys(Boc)
Test 9	Fmoc-Thr(tBu)
Test 10	Fmoc-Cys(Acm)
Test 11	Fmoc-Gly
Test 12	Fmoc-Val
Test 13	Fmoc-His(Trt)
Test 14	Fmoc-Cys(Trt)
Test 15	Fmoc-Cys(Acm)
Test 16	Fmoc-Lys(Boc)
Test 17	Fmoc-Asn(Trt)
Test 18	Fmoc-Ala
Test 19	Fmoc-Leu
Test 20	Fmoc-Ala
Test 21	Fmoc-Ser(tBu)

Test 22	Fmoc-Tyr(tBu)
Test 23	Fmoc-Leu
Test 24	Glp

(1) Access to the first protected amino acid in the main chain

Take 0.03mol of the first protected amino acid and 0.03mol of HOBt, dissolve with an appropriate amount of DMF; take another 0.03mol of DIC, slowly add it to the DMF solution of the protected amino acid under stirring, and stir and react at room temperature for 30 minutes to obtain the activated protection Amino acid solution, ready for use.

Take 0.01 mol of Rink amide MBHA resin (substitution value is about 0.3 mmol/g), use 20% PIP/DMF solution to deprotect for 25 minutes, wash and filter to obtain the resin de-Fmoc.

The activated first protected amino acid solution is added to the resin from which Fmoc has been removed, and the coupling reaction is carried out for 60-300 minutes, filtered and washed to obtain a resin containing one protected amino acid.

(2) Access to the 2nd to 29th protected amino acids in the main chain

Using the same method as the above-mentioned access to the first protected amino acid in the main chain, the above-mentioned corresponding second to 29 protected amino acids are sequentially connected to obtain a resin containing 29 amino acids in the main chain.

(3) Access to the first protected amino acid in the side chain

Use 50% HFIP/DCM solution to deprotect the side chain, repeat 5 times for 35 minutes each time, filter and wash to obtain the Mtt-deprotected resin for use.

Take 0.3mol of 2,2'-dithiodipyridine, dissolve it with an appropriate amount of DMF, add it to the above-mentioned Mtt-protected resin, stir and react for 3 hours, filter and wash to obtain SH-activated resin, which is used for later use.

Take 0.03mol of the first protected amino acid (Fmoc-Cys-Asn(Trt)-OtBu) in the side chain, dissolve it with an appropriate amount of DMF, add it to the above SH-activated resin, add 2ml DIPEA, stir and react for 3 hours, filter and wash, and complete the side. Access to the first amino acid of the chain.

(4) Access to the second protected amino acid in the side chain

Take 0.03 mol of the second protected amino acid in the side chain and 0.03 mol of HOBt, and dissolve it with an appropriate amount of DMF; take another 0.03 mol of DIC, slowly add it to the DMF solution of the protected amino acid under stirring, and stir for 30 minutes at room temperature. protected amino acid solution.

Use 20% PIP/DMF solution to deprotect for 25 minutes, wash and filter, add the second protected amino acid solution of the activated side chain to the resin from which Fmoc has been removed, and conduct coupling reaction for 60 to 300 minutes, filter and wash to obtain a side chain containing Resin that protects amino acids in the second chain.

(5) Access to the 3rd to 20th protected amino acids in the side chain

By adopting the same method as above for accessing the first protected amino acid of the main chain, the third to 20th protected amino acids corresponding to the side chain are successively connected to obtain a peptide resin.

2. Preparation of crude product

Take the above peptide resin, add a cleavage reagent with a volume ratio of TFA:Tis:water=95:5:5 (10mL of cleavage reagent/g resin), stir well, stir at room temperature for 3 hours, filter the reaction mixture with a sand core funnel, collect The filtrate and resin were washed three times with a small amount of TFA, the combined filtrates were concentrated under reduced pressure, anhydrous ether was added for precipitation, the precipitate was washed three times with anhydrous ether, and dried to obtain an off-white powder.

The obtained off-white powder was dissolved with 20% DMSO aqueous solution, adjusted to pH 7.5 with ammonia water, stirred for 10 hours, then added glacial acetic acid to 20% acetic acid, and added dropwise iodine/ethanol saturated solution under stirring until complete cyclization, 35～40 ℃ Concentrate under reduced pressure to obtain a crude concentrated solution.

3. Preparation of pure product

Take the concentrated solution of the above crude product, filter it with a 0.45 μm mixed microporous membrane, and purify it for later use;

High performance liquid chromatography was used for purification. The chromatographic packing for purification was 10 μm reversed-phase C18, the mobile phase system was 0.1% TFA/water solution-0.1% TFA/acetonitrile solution, and the flow rate of the 30mm*250mm column was 20mL/min. Gradient system elution, cyclic injection purification, take the crude product solution and load it into the chromatographic column, start the mobile phase elution, collect the main peak and evaporate the acetonitrile to obtain the purified intermediate concentrate;

The concentrated solution of the purified intermediate was filtered with a 0.45 μm filter membrane for use, and the salt was exchanged by high performance liquid chromatography. The mobile phase system was 1% acetic acid/water solution-acetonitrile, and the chromatographic packing for purification was 10 μm reversed-phase C18, 30mm*250mm The flow rate of the chromatographic column is 20mL/min (the corresponding flow rate can be adjusted according to the chromatographic column of different specifications); the gradient elution, cyclic sample loading method is adopted, the sample is loaded into the chromatographic column, the mobile phase elution is started, the spectrum is collected, and the observation Changes in absorbance, collect the main peak of changing salt and check the purity with analytical liquid phase, combine the main peak solution of changing salt, concentrate under reduced pressure to obtain pure acetic acid aqueous solution, freeze-dry to obtain 3.9 g of pure product, the purity is 98.0%, and the total yield is 5.7%. The molecular weight was 6806.1 (100% M+H).

Example 2 Preparation of Compound 2

The preparation method is the same as in Example 1, and the protected amino acids used are as follows:

4.2 g of pure product were obtained, the purity was 98.7%, and the total yield was 6.4%. The molecular weight was 6558.9 (100% M+H).

Example 3 Preparation of Compound 3

(1) The preparation method is the same as in Example 1 except that the preparation method is different in connection with 1.

(2) Access to the first protected amino acid in the side chain

Take 0.03 mol of the first protected amino acid and 0.03 mol of HOBt in the side chain, and dissolve it with an appropriate amount of DMF; take another 0.03 mol of DIC, slowly add it to the DMF solution of the protected amino acid under stirring, and stir and react at room temperature for 30 minutes to obtain the activated protected amino acid solution.

Use 50% HFIP/DCM solution to deprotect the side chain, repeat 5 times, 35 minutes each time, wash and filter, add the first protected amino acid solution of the activated side chain to the resin that has been deprotected by Mtt, and the coupling reaction is 60 ~300 minutes, filter and wash to obtain a resin containing the first protected amino acid in the side chain.

The protected amino acids used are as follows:

5.0 g of pure product was obtained with a purity of 97.5% and a total yield of 7.4%. The molecular weight was 6785.1 (100% M+H).

Example 4 Preparation of Compound 4

The preparation method is the same as in Example 3, and the protected amino acids used are as follows:

6.3 g of pure product were obtained, the purity was 97.1%, and the total yield was 9.6%. The molecular weight was 6537.8 (100% M+H).

Example 5 Preparation of compound 5

(1) The preparation method is the same as in Example 1 except that the preparation method is different in connection with 1.

(2) Access to the first protected amino acid in the side chain

Use 50% HFIP/DCM solution to deprotect the side chain, repeat 5 times for 35 minutes each time, filter and wash to obtain the Mtt-deprotected resin for use.

Take 0.03 mol of the first protected amino acid in the side chain, dissolve it with an appropriate amount of DMF, add it to the Mtt-protected resin, stir evenly, add 0.03 mol of N-methylmorpholine and 0.03 mol of lithium chloride, and stir for 6 hours. Filter and wash to obtain a resin containing the first protected amino acid in the side chain.

The protected amino acids used are as follows:

7.5 g of pure product was obtained, the purity was 98.3%, and the total yield was 11.0%. The molecular weight was 6788.1 (100% M+H).

Example 6 Preparation of compound 6

The preparation method is the same as in Example 5. The protected amino acids used are as follows:

5.5 g of pure product was obtained, the purity was 98.7%, and the total yield was 8.4%. The molecular weight was 6540.9 (100% M+H).

Example 7 Preparation of compound 7

(1) The preparation method is the same as in Example 1 except that the preparation method is different in connection with 1.

(2) Access to the first protected amino acid in the side chain

Take 0.03 mol of the first protected amino acid and 0.03 mol of HOBt in the side chain, and dissolve it with an appropriate amount of DMF; take another 0.03 mol of DIC, slowly add it to the DMF solution of the protected amino acid under stirring, and stir and react at room temperature for 30 minutes to obtain the activated protected amino acid solution

Use 2% hydrazine hydrate/DMF solution to deprotect the side chain, repeat 3 times for 10 minutes each time, wash and filter, add the first protected amino acid solution of the activated side chain to the resin that has been de-Dde removed, and the coupling reaction is performed for 60 ~300 minutes, filter and wash to obtain a resin containing the first protected amino acid in the side chain.

The protected amino acids used are as follows:

8.2 g of pure product were obtained, the purity was 98.1%, and the total yield was 12.1%. The molecular weight was 6770.1 (100% M+H).

Example 8 Preparation of Compound 8

The preparation method is the same as in Example 7. The protected amino acids used are as follows:

7.6 g of pure product were obtained, the purity was 97.9%, and the total yield was 11.7%. The molecular weight was 6522.9 (100% M+H).

Example 9 Determination of preliminary pharmacokinetic properties

Each compound was divided into two administration groups: SD rats, 4 males in each group, 8 rats in total.

Intravenous tail vein group: the dose was 1 mg/kg, blood was collected from the orbital vein of the rats before the drug (0h) and 30min, 1h, 2h, 4h, 8h, 24h, 48h, 96h, and 144h after the drug, and centrifuged. Plasma sample.

Subcutaneous administration group: the dose was 1 mg/kg, blood was collected from the orbital vein of rats before the drug (0h) and 1h, 2h, 3h, 4h, 8h, 24h, 48h, 96h, 144h after administration, and the plasma was separated by centrifugation. sample.

The plasma concentrations of the corresponding compounds in the plasma samples of SD rats were determined by LC-MS method. After intravenous and subcutaneous administration, the half-life of compound SD rats administered subcutaneously (SC) is shown in the following table:

compound	t 1/2 (h)
Compound 1	8.6
Compound 2	11.2
Compound 3	8.2
Compound 4	9.9
Compound 5	9.0
Compound 6	10.3
Compound 7	10.5
Compound 8	12.9

The long-acting relaxin 2 analog provided by the present invention and its use are described in detail above. The principles and implementations of the present invention are described herein by using specific examples, and the descriptions of the above embodiments are only used to help understand the method and the core idea of the present invention. It should be pointed out that for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can also be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims (6)

1. A long-acting relaxin 2 analog having the structure of formula I:

   

   When X1 in formula I is S, X2 is S, or is CH2;

   When X1 in formula I is CH2, X2 is S, or is CH2;

   X1 in formula I is NH, and X2 is CO;

   When X1 in formula I is CO, X2 is NH;

   AA1 in formula I is any codable amino acid except Cys and Glu, or any non-codable amino acid without SH group;

   AA2 in formula I is Asp, or Asn, or Glu, or GIn, or GIn, or Ser, or Thr;

   AA3 in formula I is any codable amino acid except Cys, or any non-codable amino acid without SH group, or is absent;

   AA4 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

   AA5 in formula I is any codable amino acid except Cys, or any non-codable amino acid without SH group, or is absent;

   AA6 in formula I is any codable amino acid except Cys, or any non-codable amino acid without SH group, or is absent;

   AA7 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

   AA8 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

   AA9 in formula I is any codable amino acid except Cys, or is any non-codable amino acid without SH group, or is absent;

   AA10 in formula I is Lys, or Dah, or Orn, or Dab, or Dap;

   AA11 in formula I is NH2, or OH;

   R in formula I is cholesterol succinate monoester, or 2-cholesteryl acetic acid, or 2-cholesteryl propionic acid, or 3-cholesteryl propionic acid, or 2-cholesteryl butyric acid, or 2-cholesteryl isobutyrate acid, or 3-cholesterylbutyric acid, or 3-cholesterylisobutyric acid, 4-cholesterylbutyric acid, or 2-cholesterylvaleric acid, or 2-cholesterylisovaleric acid, or 3-cholesterylvaleric acid, Either 5-cholesteryl valeric acid, or 2-cholesteryl caproic acid, or 6-cholesteryl caproic acid, or 2-cholesteryl heptanoic acid, or 7-cholesteryl heptanoic acid, or 2-cholesteryl caproic acid, or 8 -Cholesterol octanoic acid, or HO2C(CH2)n1CO-(γGlu)n2-(PEGn3( _CH2 )n4CO)n5-;

   Where: n1 is an integer from 10 to 20;

   n2 is an integer from 1 to 5;

   n3 is an integer from 1 to 30;

   n4 is an integer from 1 to 5;

   n5 is an integer from 1 to 5;

   When AA11 in formula I is OH, X1 is S, and X2 is CH2, R can be absent;

   When AA11 in formula I is OH, X1 is CH2, and X2 is S, R can be absent;

   When AA11 in formula I is OH, X1 is CH2, and X2 is CH2, R can be absent;

   When AA11 in formula I is OH, X1 is NH, and X2 is CO, R can be absent;

   When AA11 in formula I is OH, X1 is CO, and X2 is NH, R can be absent;

   When AA11 in formula I is NH2, X1 is S, and X2 is S, R can be absent;

   When AA11 in formula I is NH2, X1 is S, and X2 is CH2, R can be absent;

   When AA11 in formula I is NH2, X1 is CH2, and X2 is S, R can be absent;

   When AA11 in formula I is NH2, X1 is CH2, and X2 is CH2, R can be absent;

   When AA11 in formula I is NH2, X1 is NH, and X2 is CO, R can be absent;

   When AA11 in formula I is NH2, X1 is CO, and X2 is NH, R may be absent.
2. A kind of long-acting relaxin 2 analog according to claim 1, is characterized in that, comprises the pharmaceutically acceptable salt, solvate, chelate or non-covalent complex formed by the analog, based on the A prodrug on a compound basis, or any mixture of the above.
3. The application of a relaxin 2 analog according to claim 1 or 2 in the preparation of a pharmaceutical composition for treating diseases.
4. Use of the pharmaceutical composition according to claim 3 for treating various diseases, including various types of heart failure and various inflammatory conditions.
5. The use according to claim 4, wherein the various types of heart failure include acute congestive heart failure, compensated heart failure and the like.
6. The use according to claim 4, wherein the various inflammatory conditions mainly include eosinophilic airway hyperresponsiveness, asthma, rheumatoid arthritis, multiple sclerosis, ankylosing spondylitis, and inflammatory bowel disease , gout, myositis, systemic lupus erythematosus, vasculitis, sepsis, trauma, wound healing, eczema, dermatitis, scleroderma, acne, urticaria, psoriasis and allergic reactions.