WO2014005513A1

WO2014005513A1 - Cyclopeptide lhrh antagonist derivatives and pharmacal use thereof

Info

Publication number: WO2014005513A1
Application number: PCT/CN2013/078713
Authority: WO
Inventors: 刘克良; 周宁; 李改桃; 吕玉健; 冯思良; 周文霞; 张永祥; 程军平; 郄建坤; 梁远军; 许笑宇; 王晨宏; 孟庆斌; 韩寒; 徐亮; 蔡俐锋
Original assignee: 中国人民解放军军事医学科学院毒物药物研究所
Priority date: 2012-07-05
Filing date: 2013-07-03
Publication date: 2014-01-09
Also published as: CN103524599B; CN103524599A

Abstract

Disclosed in the present invention are cyclopeptide derivatives with LHRH acceptor antagonistic activity of general formula I, the preparation method thereof, the pharmaceutical composition containing them and uses thereof in the preparation of medicines for treating prostate cancer, endometrial cancer, ovarian cancer, breast cancer and other sexual hormone dependent diseases related to reproduction, and of contraceptives.

Description

Cyclopeptide LHRH antagonist derivative and its pharmaceutical use

The present invention relates to a cyclic peptide derivative having a luteinizing hormone releasing hormone (LHRH) receptor antagonistic activity, having an action of inhibiting pituitary secretion of gonadotropin and inhibiting gonadal secretion hormone, a preparation method thereof, a pharmaceutical composition containing the same, and They are used in the preparation of medicaments for the treatment of prostate cancer, endometrial cancer, ovarian cancer, breast cancer and other reproductive-related sex hormone-dependent diseases, contraception and the like. Background technique

LHRH, also known as GnRH (tropin releasing hormone), its main physiological function is to regulate the body's sex hormones. LHRH in the body is mainly secreted and secreted by hypothalamic (Hypothalamus) neurons, released to the pituitary ry portal veins at a pulse frequency of about one hour, by binding to specific LHRH receptors on the pituitary. , the pituitary gland to secrete luteinizing hormone (LH) and follicle stimulating hormone (FSH) two gonadotropins. LH and FSH reach the male gonadal testis (Testis) with blood circulation to promote spermatogenesis (Spermatogenesis) and testosterone (T) synthesis; or reach the female gonadal ovary (Ovary) to promote follicle formation ( Follictilogenesis) and estrogen (Estrone, E) and progesterone (P) synthesis. Finally, the steroidal sex hormones T, E and P are fed back to the hypothalamus and pituitary. Thus, it can be seen that LHRH plays a key role in the regulation of the physiological function of the sex hormones in the hypothalamus.

LHRH consists of ten amino acid residues and the C-terminus contains an amide structure. The primary structure of LHRH is as follows: p-Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH ₂

LHRH antagonists can inhibit the release of LH by blocking the action of LHRH, and thus can be used for the treatment of sex hormone related diseases such as prostate cancer. Compared with agonists, LHRH antagonists have a rapid onset, no overshoot, rapid recovery after withdrawal, and serum androgen water. The advantages of high controllability and so on. It is expected that LHRH antagonists are superior to agonists in the treatment of prostate cancer, are more acceptable to patients, and have greater application prospects than agonists.

Although there are many LHRH antagonists currently developed, as a peptide drug, most of them still have shortcomings such as low bioavailability, short half-life in vivo, and high histamine release. Similar to other peptide drugs, LHRH antagonist drugs are also difficult to absorb by oral administration. To the best of our knowledge, currently marketed LHRH antagonist drugs are administered parenterally. Compared with linear peptides, cyclic peptides have better enzyme stability, and some cyclic peptides can enhance oral absorption capacity because of enhanced lipophilicity. Therefore, the object of the present invention is to convert linear LHRH decapeptide antagonist derivatives into The cyclic peptide is expected to improve its enzyme stability and membrane permeation ability while maintaining its original activity, thereby obtaining a novel or even oral LHRH antagonist drug. Summary of the invention

The present inventors have discovered that a cyclic peptide derivative of formula I has been discovered.

Ac-D-Nal -D-Cpa ² -D-Xaa ³ -Xaa -Xaa ⁵ -Xaa ⁶ -Leu ⁷ -Xaa ⁸ -Y-NH ₂

I

Or a stereoisomer thereof or a physiologically toxic salt thereof, which has good LHRH antagonistic activity and high enzyme stability, and thus can be used as a medicament for prostate cancer, endometrial cancer, ovarian cancer, breast cancer and other sex hormones. Depends on the use of the relevant disease treatment.

Thus the first aspect of the invention relates to a compound of formula I, or a stereoisomer thereof or a salt of no physiological toxicity,

Ac-D-Nal ¹ -D-Cpa ² -D-Xaa ³ -Xaa -Xaa ⁵ -Xaa ⁶ -Leu ⁷ -Xaa ⁸ -Y-NH _:

I

among them,

Xaa ³ is D-Phe or D-Pal;

Xaa ⁴ is L or D type Cys, Asp, Glu or Lys; Xaa ⁵ is L or D type Aph (Hor) or Arg;

Xaa ⁶ is L or D type Aph (Cbm) or Pal;

Xaa ⁸ is Ilys or Arg;

Y is L or D-type Cys-Ala, Asp-Ala, GIu-Ala, Lys-Ala, Pro-Cys, Pro-Asp, Pro-Glu or Pro-Lys, wherein two amino acids may each independently be L-form or Type D;

Among them, the Cys side chain thiol group of 4 and Y can be ring-formed by two bowls, or the side chain of 4th and Y amino acids Asp, Glu, Lys and the like can be formed by an amide bond.

A compound according to any one of the first aspects of the invention, which is selected from one or more of the following (1) to (5):

(1) Xaa ³ is D-Pal;

(2) Xaa ⁴ is Cys;

(3) Xaa ⁵ is D-Aph (Hor);

(4) Xaa ⁶ is D-Aph (Cbm);

(5) Y is Cys-D-Ala or Pro-D-Cys.

A compound according to any one of the first aspects of the invention, wherein Xaa ³ is D-Pal.

A compound according to any one of the first aspects of the invention, wherein Xaa ⁴ is Cys.

A compound according to any one of the first aspects of the invention, wherein Xaa ⁵ is D-Aph (Hor). A compound according to any one of the first aspects of the invention, wherein Xaa ⁶ is D-Aph (Cbm). The compound according to any of the first aspects of the invention, wherein Y is Cys-D-Ala or Pro-D-Cys.

A compound according to any one of the first aspects of the invention, which is selected from the group consisting of:

1), Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys"-Aph(Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Cys ⁹ -D-Ala ¹⁰ -NH ₂ -

2), Ac-D-Nal ¹ - D-CpaS-D-PaP-Cy Is ⁴ - Aph(Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -DC Iys ⁹ -D-Ala ^{, 0} -NH ₂ ;

3), Ac-D-Nal-D-Cpa ² -D-Pal ³ -Cys ⁴ -Aph(Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Pro ⁹ -Cys ¹⁰ -NH ₂ ;

4), Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys -Ap (Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Pro ⁹ -D-Cys ¹⁰ - NH ₂ ; 5), Ac-D-Nal'-D-Cpa ² -D-Phe -Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu ⁷ -Arg ⁸ -Pro ⁹ -Cys ^I0 -NH ₂ .

6), Ac-D-Nal ¹ -D-Cpa ² -D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal -Leu ⁷ -Arg ⁸ -Cys ⁹ -D-Ala ^{, 0} -NH ₂

7), Ac-D-Nal '-D-Cpa^D-Phe^Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu B Arg ⁸ -Pro ⁹ -D-Cys ¹⁰ -NTH ₂ is a compound 1, 2 3, 4, 5, 6, and 7.

Preferably, the compound of the invention is selected from the group consisting of compounds 2, 3, 4, 5, and a second aspect of the invention relates to a pharmaceutical composition comprising at least one compound of any of the first aspects of the invention, and optionally a pharmaceutically acceptable drug Use excipients, carriers or excipients.

A third aspect of the invention relates to the use of a compound according to any of the first aspects of the invention for the manufacture of a medicament for the prevention and/or treatment of a sex hormone related disease or for contraception.

The use according to the third aspect of the invention, wherein the sex hormone related diseases include sex hormone related cancer, adolescent precocious puberty, benign prostatic hyperplasia, endometriosis, irregular menstruation (such as amenorrhea, dysfunctional uterine bleeding or ovulation disorder) or Infertility and so on.

The use according to the third aspect of the invention, wherein the sex hormone related cancer comprises prostate cancer, endometrial cancer, breast cancer or ovarian cancer or the like. A fourth aspect of the invention relates to a compound according to any one of the first aspects of the invention for use in the preparation of a gonadotropin-secreting hormone (e.g., LH and/or FSH) and/or a gonadotropin-secreting hormone (e.g. estrogen, pregnancy) Use of drugs for hormones and/or testosterone). A fifth aspect of the invention relates to a process for the preparation of a compound according to any one of the first aspects of the invention, characterized in that:

Using MBHA resin as carrier, Boc- or Fmoc-protection strategy, DCC/HOBT or BOP/DIEA as condensation reagent, HC1/dioxane or piperidine/DMF as deprotection reagent, after amino acid condensation is completed, use 1 ₂ off In addition to the Trt protecting group on the thiol group, the two sulfhydryl groups in the molecule are oxidized by a disulfide bond, and after the reaction, the cyclic peptide derivative is cleaved from the MBHA resin by liquid HF. A sixth aspect of the invention relates to a method of preventing and/or treating a sex hormone related disease or contraception, the method comprising the step of preventing or treating an effective amount of a compound of any one of the first aspects of the invention to a subject in need thereof.

The method according to the sixth aspect of the present invention, wherein the sex hormone related diseases include sex hormone related cancer, adolescent precocious puberty, benign prostatic hyperplasia, endometriosis, irregular menstruation (such as amenorrhea, dysfunctional uterine bleeding or ovulation disorder) or Infertility and so on.

According to the method of the sixth aspect of the invention, the sex hormone-related cancers include prostate cancer, endometrial cancer, breast cancer or ovarian cancer, and the like. A seventh aspect of the invention relates to a method of inhibiting pituitary secretion of gonadotropins (e.g., LH and/or FSH) and/or inhibiting gonadal secretory hormones (e.g., estrogen, progestin, and/or testosterone), the method comprising The subject is in need of an effective amount of a compound of any of the first aspects of the invention. Unless otherwise stated, the terms used in this application have the following meanings.

In the present invention, all amino acid configurations are L-form except for the D-type.

In the present invention, the disulfide bond refers to a -S-S formed between a cysteine and a cysteine side chain.

In the present invention, the amide bond refers to -CO-NH - formed between an amino acid and an amino acid. According to the present invention, the cyclic peptide derivative represented by Formula I, and its stereoisomers and its non-physiologically toxic salt exhibit not only good effects in animal antiandrogen secretion experiments, but also exhibit higher enzymes in in vitro assays. It is stable and lipophilic and can therefore be used as a hormonal agent in animals, preferably in mammals, especially humans.

The invention therefore also relates to a pharmaceutical combination comprising at least one cyclic peptide derivative of the formula I and/or a stereoisomer thereof or a non-physiologically toxic salt thereof, and a conventional pharmaceutical excipient or adjuvant, in an effective amount as an active ingredient. Things. "Conventional pharmaceutical excipients or adjuvants" herein includes any or all solvents, dispersion media, coatings, antibacterial or antifungal agents, isotonic and sustained release agents, And similar physiologically compatible preparations for intravenous, intramuscular, subcutaneous, or other modes of administration, such as oral administration. Depending on the mode of administration, the active compound may be coated to protect the compound from the effects of acid or other natural conditions.

The term "physiologically toxic salt" as used in the present invention means a salt or a combination thereof which retains the expected physiological activity of the parent compound without causing any unexpected toxic side effects. For example: hydrochloride, hydrobromide, sulfate, phosphate, nitrate, and acetate, oxalate, tartrate, succinate, phthalate, benzoate, pamoate Salt, alginate, methanesulfonate, naphthalenesulfonate, etc. The cation according to the salt may be: an inorganic salt such as a potassium salt, a lithium salt, a zinc salt, a copper salt, a cerium salt, a cerium salt or a calcium salt, or an organic salt such as a trialkylammonium salt.

The cyclic peptide derivative of the formula (1) and a stereoisomer thereof or a pharmaceutical composition containing the same can be administered in any manner known per se, such as oral, intramuscular, subcutaneous, etc., in a dosage form such as a tablet or a capsule. , buccal tablets, chewable tablets, tinctures, suspensions, transdermal agents, micro-emulsions, implants, syrups, and the like. It may be a general preparation, a sustained release preparation, a controlled release preparation, and various microparticle delivery systems. In order to form a unit dosage form into tablets, various biodegradable or biocompatible carriers well known in the art can be widely used. Examples of the carrier are, for example, a saline base and various buffered aqueous solutions, ethanol or other polyols, liposomes, polylactic acid, vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and the like.

The dose of the cyclic peptide derivative represented by Formula I of the present invention or a stereoisomer thereof depends on many factors such as the nature and severity of the disease to be prevented or treated, the sex, age, body weight, sensitivity of the patient or animal. And the individual reaction, the specific compound used, the route of administration, the number of administrations, and the desired therapeutic effect. The above dosages may be administered in a single dosage form or divided into several, for example two, three or four dosage forms. The single maximum dose is generally not more than

30 mg/kg body weight, for example 0.001-30 mg/kg, preferably 0.01-5 mg/kg, preferably a dose range of 0.5-2 mg/kg body weight. However, in some cases, it is also possible to use a single dose of 30 mg/kg or more or 0.001 mg/kg or less. detailed description

The abbreviations used in the present invention have the following meanings: Ac-acetyl

Ala - propoxylate,

Arg-arginine,

Aph 4-aminophenylalanine

Hor - whey acyl

Cbm-carbamoyl

Boc-tert-butoxycarbonyl,

BOP-benzotrix-1-oxo-tris(dimethylamino)phosphorus hexafluorophosphate,

Cys-cysteine

DCC-dicyclohexylcarbodiimide,

DIEA - diisopropylethylamine,

HOBt- 1-hydroxybenzotriazole,

Leu - leucine,

ILys-isopropyl lysine

MBHA- phenylamino hydrazine based resin,

Nal-naphthylalanine,

Pal -3-pyridine alanine,

Phe- phenylalanine,

RP-HPLC - Reversed Phase High Performance Liquid Chromatography

Pro - proline,

Ser-serine,

TEA - triethylamine,

C _P a-4-chlorophenylalanine

The embodiments of the present invention are described in detail below with reference to the accompanying drawings. If no specific conditions are specified in the examples, they are carried out according to the general conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained commercially. The solid phase synthesis carrier MBHA resin used in the examples is Tianjin Nankai Synthetic Co., Ltd.; DCC, HOBT BOP, DIEA and Boc-protected or Ftnoc-protected natural amino acids are produced by Shanghai Jill Biochemical Co., Ltd. and Chengdu Kaitai New Technology Co., Ltd. , Boc- or Fmoc-protected non-natural amino acids are provided by our laboratory, except as indicated. Example 1 : Synthesis of a cyclic peptide LHRH antagonist derivative

Weigh 0.5 g of MBHA resin into the reactor, add DCM (4-dicyanomethylene) to swell for 30 min, and stir magnetically to uniformly disperse the resin. Drain, wash with DCM, sterol (MeOH), DCM (3x2 min) (the same below), and drain. Neutralize (2 x 5 min) with 10% DIEA/DCM, free amino groups, wash the resin, and drain. (1) Add 1.08 mmol of Fmoc-D-Ala-OH, 1.08 mmol HOBt, 1.08 mmol DIC (N, N,-diisopropylcarbodiimide) in 2 mL CH ₃ OH, 4 mL DCM at room temperature 4 b. (2) Wash, pump a thousand, take a little resin and test with ninhydrin indicator. A small amount of resin was taken in a small glass test tube, and two drops of three solutions of ninhydrin were added in sequence. Heated at 110 C for 5 min. Positive: Most resins are blue or light red, solution Blue; Negative: The resin is colorless and transparent, and the solution is light yellow in color. If it is positive, repeat (1). (3) If it is negative, 10 amino acids are sequentially coupled to the resin in the order from the C-terminus to the N-terminus.

(4) A: When all the amino acids have been condensed, 10 times the amount of 1 ₂ is added to the peptide resin, and DMF (dimethylformamide) is used as a solvent, and reacted at room temperature for 4 h to form a ring. After the reaction is completed, the protective group and the resin are cleaved by HF cleavage, and lyophilized to obtain a crude peptide. The pure peptide was obtained by medium pressure chromatography and HPLC analysis. Compound 5, 6, 7, a compound was synthesized by this method.

5), Ac-D-Nal'-D-Cpa ² -D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu ⁷ -Arg ⁸ -Pro ⁹ -Cys ^{, 0} -NH ₂

6), Ac-D-Nal ¹ -D-Cpa ² -D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu ⁷ -Arg ⁸ -Cys ⁹ -D-Ala ^l0 -NH ₂

7), Ac-D-Nal ' -D-Cpa ² - D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal -Leu ⁷ -Arg ⁸ -Pro ⁹ -D-Cys ¹⁰ -NH ₂

(4) B: After all the amino acids have been condensed, the Boc protecting group is removed with 25% TFA, washed, and 1.08 mmol of Hor, 1.08 mmol of HOBt, 1.08 mmol of DlC is added, and the reaction is carried out for 4 h at room temperature. Detection, if positive, re-react until negative. Then, 10 times the amount of ruthenium _{2 was added} , and DMF was used as a solvent, and reacted at room temperature for 4 h to form a ring. After completion of the reaction, the protective group and the resin were cleaved by HF cleavage, and lyophilized to obtain a crude peptide. The pure peptide was obtained by medium pressure chromatography and HPLC analysis. In this way, compounds 1, 2, 3, 4, compounds are synthesized.

\ ) , Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys"-Ap (Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Cys ⁹ -D-Ala ^{1 Q} -NH ₂

), Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys -Aph(Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -D-Cys ⁹ -D-Ala ¹⁰ -NH _:

3), Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys -Aph(Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Pro ⁹ -Cys ¹⁰ -NH _;

4), Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys"-Aph{Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -ltys ⁸ -Pro ⁹ -D-Cys ¹⁰ The MS/HPLC data of each compound synthesized above -NH are shown in Table 1. Table 1 MS/HPLC data of synthetic cyclic peptides

Compound MS. _Bv . MS _cal , Purity

5 1460.2 1460.3 >99%

6 1434.0 1434.1 96.62 7 J460.2 1460.3 >99%

1 1652.2 1652.3 >98%

2 1652.2 1652.3 >95%

3 1678.8 1678.4 >95%

4 1678.3 1678.4 >90% Example 2: Inhibition of testosterone in rats

Before the experiment, the weight of the animals (SD male rats) was measured, and the blood was collected from the posterior venous plexus of the glass capillary. After separating the serum, the serum testosterone content was determined by chemiluminescence method. The testosterone content and body weight were randomly grouped, and 4 animals in each group were used in one time. Excessive subcutaneous injection of test compound 500

After 8 hours, 16 hours, 24 hours, etc., blood was collected from the posterior venous plexus, centrifuged for 8 min at SOOOrpm, and the serum obtained was determined by chemiluminescence (American Beckman Coulter Access Immunoassay System chemiluminescence). The specific knots are shown in Table 2. It can be seen that the compounds of the present invention are effective in inhibiting testosterone activity in rats. Example 3: Determination of antagonistic activity against GnRH (gonadotropin-releasing hormone) receptor GnRH receptor function detection using a calcium-sensitive fluorescent dye, which can initiate initiation of the endoplasmic reticulum when the GnRH receptor is activated by an agonist Calcium ion release, calcium ions can bind to fluorescent dyes and enhance the fluorescence intensity. The stronger the fluorescent signal, the stronger the activity of the ligand or agonist, and the weaker the activity of the ligand or agonist. Since the antagonistic compound can compete with the GnRH receptor, the release of calcium ions in the endoplasmic reticulum is reduced, and the fluorescent signal is also reduced. The inhibitory activity of the compound to be detected on the GnRH receptor is characterized in terms of the degree of signal reduction.

CHO-Kl/Gal5 cells stably expressing the GnRH receptor (purchased from GenScript) were inoculated into 384-well plates, routinely cultured, and passaged. Fluorescent dye is added, and then the prepared test compound solution (detection concentration: le-9M; le-lOM; le-ΠΜ; le-12M; le-13M; le-14M; le-15M; le-16M) The corresponding DMSO control was 1% v/v.) Into the cell plate, after incubation, the calcium ion fluorescence detection kit (Molecular Devices, Inc.;) was used. The overall detection time of the instrument was 120 seconds, at 21 seconds. Automatically add LHRH to the cell plate for detection (Reference: Jack J. Chen et al, Identification of a new class of small molecule C5a receptor antagonists* Bioorganic & Medicinal Chemistry Letters 20 (2010) 662-664).

Data acquisition and analysis were performed using Excel and GraphPad Prism 4 software programs. For each detection well, the average fluorescence intensity value of 1 to 20 seconds is used as the baseline, and the maximum fluorescence intensity value of 21 to 120 seconds minus the baseline value is the relative fluorescence intensity value (ARFU), from which the value can be calculated. Percent inhibition. The data is processed using a four-parameter equation using GraphPad Prism 4 to calculate EC50 or IC50 values. The specific results are shown in Table 2. It can be seen that the compound of the present invention has an antagonistic activity against the GnRH receptor. Example 4: Determination of oil-water partition coefficient

Weigh about 0.5 mg of the sample (test compound), dissolve it in 0.5 mL of water, centrifuge at high speed, and take the supernatant for HPLC detection. Determine the compound in different ratios of methanol/phosphate buffer (pH=7.4) by RP-HPLC. The retention time t _R (i.e., the retention time in Table 2) was measured using a NaN0 ₂ solution. , Calculate the capacity factor k = (t _R -t.) / t. . Taking the methanol content as the abscissa and the logk as the ordinate as the curve, the longitudinal moment logD is obtained. _Ct (equivalent to the logk of the compound at a methanol content of 0) is the oil-water partition coefficient value (see F. Lombardo, MY Shalaeva, KA Tupper, et al. ElogPoct: a tool for lipophilicity determination in drug discovery. Journal of medicinal chemistry > 2000, 43 ( 15 ) : 2922 - 2928; F. Lombardo, MY Shalaeva, KA Tupper, et al. ElogDoct: A tool for lipophilicity determination in drug discovery. 2. Basic and neutral compounds [J]. Journal of medicinal chemistry , 2001, 44 ( 15 ) : 2490 - 2497 ). The specific results are shown in Table 2. It can be seen that the cyclic peptide compound of the present invention has higher lipophilicity (longer retention time) than Degardix. Example 5: Stability evaluation: Determination of half-life in artificial intestinal fluid

Mix the aqueous test compound (0.2 mg/mL) with artificial intestinal juice (see Qian Zhongzhi, Qi Ping, Wang Guorong. Chinese Pharmacopoeia, Appendix II, Chinese Drug Standard, 2005. 158), and incubate in a 37 °C constant temperature oscillator. , take out at different time points (0, 10, 20, 40, 60, 90, 120 min), stop the reaction with 10% trifluoroacetic acid (TFA); high speed centrifugation (10000 r/min) 10 min, the supernatant was taken for HPLC analysis, and the concentration was calculated from the peak area. Taking the time t as the abscissa, the common logarithm (logc) of the substrate concentration is plotted on the ordinate, and logc = -at + b, and the half-life t _1/2 = 0.3 / a. The specific results are shown in Table 2. It can be seen that the compounds of the present invention have high enzyme stability. Table 2

Compound No. GnRH receptor antagonistic activity half-life in rats (human oil moisture retention time testosterone concentration a sex IC50 (mol L ) process intestinal fluid) distribution coefficient (mm)

Degarelix completely castrated 1.7181e-012 4 (U h 4.5058 3.67

1 Not castrated 6.1879e-009 58.2 h 4.3745 4.43

2 Complete castration 1.2888e-011 50.4 h 6.113 3.80

3 Complete castration 3.9485Θ-012 61.3 h 5.7201 4.42

4 Partial castration 1.1967e-010 47.5 h 5.9224 4.43

5 Complete castration Not determined 29.4 h 3.6262 12.22

6 Complete castration Not determined 11.4 h 3.6149 9.77

7 Complete castration was not determined 0.25 h 3.4674 11.36 a: Subcutaneous administration, 500 yg/kg, testosterone concentration after 8 hours. Inhibition to ln _g /mL or less is complete castration. Less than the testosterone concentration of the control group, but not to l ng/mL for partial castration;

b: Determined in 77% methanol/stone solution buffer (7, 4). Aglicntl200, 4.6*150mm SB-C18 column.

Degare l i x is a commercial gonadotropin releasing hormone for prostate cancer hormone therapy.

(GnRH) blocker, which can cause a continuous decrease in testosterone levels.

Although specific embodiments of the invention have been described in detail, those skilled in the art will understand. Various modifications and alterations may be made to those details in accordance with the teachings of the invention, which are within the scope of the invention. The full scope of the invention is indicated by the appended claims and any equivalents thereof.

Claims

Rights request

1. A compound of the formula I, or a stereoisomer thereof or a salt of no physiological toxicity,

Ac-D-Nal ¹ -D-Cpa ² -D-Xaa ³ -Xaa -Xaa ⁵ -Xaa ⁶ -Leu ⁷ -Xaa ⁸ -Y-NH ₂

I

among them,

Xaa ³ is D-Phe or D-Pal;

Xaa ⁴ is L or t> type Cys, Asp, Glu or Lys;

Xaa ⁵ is L or D type Aph (Hor) or Arg;

Xaa ⁶ is L or D type Aph (Cbm) or Pal;

Xaa ⁸ is Ilys or Arg;

Y is L or D type Cys-Ala, Asp-Ala, Glu-Ala, Lys-Ala, Pro-Cys, Pro-Asp, Pro-Glu or Pro-Lys, wherein the two amino acids may each independently be L-form or Type D;

Among them, the Cys side chain thiol group of 4 and Y may be ring-formed by a disulfide bond, or the side chain of the 4th and Y amino acids Asp, Glu, Lys or the like may be formed by an amide bond.

2. A compound according to claim 1 which is selected from one or more of the following (1) to (5):

(1) Xaa ³ is D-Pal;

(2) Xaa ⁴ is Cys;

(3) Xaa ⁵ is D-Aph (Hor);

(4) Xaa ⁶ is D-Aph (Cbm);

(5) Y is Cys-D-Ala or Pro-D-Cys. 3. A compound according to claim 1 or 2 selected from the group consisting of:

1), Ac-D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys -Aph(Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Cys ⁹ -D-Ala ⁰ - NH

2), Ac-D-Nal^D-Cpa^D-Pa-Cys^AphiHor^-D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -D-Cys ⁹ -D-Ala ¹⁰ -NH ₍

3), -D-Nal ¹ -D-Cpa ² -D-Pal ³ -Cys -Ap (Hor) ⁵ -D-Aph (Cbm) ⁶ -Leu ⁷ -llys ⁸ -Pro ⁹ -Cys ¹⁰ -NH

4),

5 ) , Ac-D-Nal ¹ -D-Cpa ² -D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu ⁷ -Arg ⁸ -Pro ⁹ -Cys ^{, 0} -NH ₂ .

6), Ac-D-Nal ¹ -D-Cpa ² -D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu ⁷ -Arg ⁸ -Cys ⁹ -D-Ala ^{l 0} -NH ₂ ·

7), Ac-D-Nal ¹ -D-Cpa ² -D-Phe ³ -Cys ⁴ - Arg ⁵ - D-Pal ⁶ -Leu ⁷ -Arg ⁸ -PiO ⁹ -D-Cys ^{, 0} -NH ₂ .

4. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 3, and optionally a pharmaceutically acceptable adjuvant, carrier or excipient.

Use of a compound according to any one of claims 1 to 3 for the preparation of a medicament for the prevention and/or treatment of a sex hormone related disease or for contraception.

6. The use of claim 5, wherein the sex hormone related diseases include sex hormone related cancers (e.g., prostate cancer, endometrial cancer, breast cancer or ovarian cancer), puberty precocious puberty, benign prostatic hyperplasia, endometriosis, menstruation Tune (such as amenorrhea, dysfunctional uterine bleeding or ovulation disorders) or infertility.

7. A compound according to any one of claims 1 to 3 for use in the preparation of a gonadotropin-secreting hormone (e.g., LH and/or FSH) and/or a gonadotropin-secreting hormone (e.g., estrogen, progestin, and/or testosterone). ) the use of the drug.

A process for the preparation of a compound according to any one of claims 1 to 3, characterized in that: MBHA resin is used as a carrier, Boc- or Fmoc-protection strategy, DCC/HOBT or BOP/DIEA is a condensation reagent, HC1/dioxane after piperazine ring or a sneeze / DMF as deprotection agent, after completion of the condensation of amino acids, with ₁₂ the removal of the protecting group Trt mercapto group, a mercapto group while two intramolecular disulfide oxidation of a ring, the reaction was complete liquid HF The cyclic peptide derivative was cleaved from the BHA resin.

A method for preventing and/or treating a sex hormone related disease or contraception, the method comprising the step of preventing or treating an effective amount of the compound according to any one of claims 1 to 3 to a subject in need thereof.

10. The method of claim 9, wherein the sex hormone related diseases include sex hormone related cancers (e.g., prostate cancer, endometrial cancer, breast cancer or ovarian cancer), puberty precocious puberty, benign prostatic hyperplasia, endometriosis, menstruation Tune (such as amenorrhea, dysfunctional uterine bleeding or ovulation disorders) or infertility.

11. A method of inhibiting secretion of a gonadotropin (e.g., LH and/or FSH) from a pituitary gland and/or inhibiting a hormone secreting hormone (e.g., estrogen, progestin, and/or testosterone), the method comprising administering to a subject in need thereof An effective amount of the step of any one of claims 1 to 3.