TW379216B - Novel LH-RH antagonists having improved action, harmaceutical composition comprising the same and process for preparing the same - Google Patents

Description

Niu C: Revised page of the Chinese specification of Patent Application No. 85114601 '1 Supplement I Min_June_5. Invention Description (9) (Please read the precautions on the back before filling this page) Synthesis of the compound of formula (V) Or it can be carried out by traditional fragmentation method or by solid phase synthesis method according to Merrifield, using D-lysine acids whose side chains have been converted with carboxylic acids of general formula (VII), and they can be established one by one in turn. -Lysine acid ° Amido linkage in the side chain, reacting the decapeptide unit with the appropriate residual acid. Thus, according to the present invention, there are three alternative methods for preparing compounds of the general formula (V). The first may include the steps (a) having an appropriate protecting group for the α-amino group and the carboxylic acid group of D-lysine acid or D-ornithine, and (b) the D-lysine acid or D having a protecting group —Ornithine reacts with a carboxylic acid of the general formula (VII) R 4-C Ο Ο Η (VII) wherein R 4 is as defined above, (c) removes the α-phosphonic acid group of the compound obtained in step (b) The protective group is incorporated in position 6 in step (h). It is printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. (D) The D-alanine having a protective group on the amine group is coupled to a resin-type solid. On the support (Merrifield synthesis method), (e) remove the protective group on the amine group of alanine, (f) make the alanine bonded to the solid support and the proline acid having a protective group on the nitrogen atom Reaction, (g) removing the protecting group on the nitrogen atom of proline,

(h) Use amino acids 1 to 8 of formula (V) in the order of 8 to 1; use the modified D for position 6 described in step (c). D This paper size applies Chinese National Standard (CNS ) A4 size (210 x 297 mm) -12-A7 B7 V. Description of the invention (1) The present invention relates to a novel LH-RH antagonist, especially a peptidomimetic and a peptide with modified side chains. Salts formed from acceptable acids, and a method for preparing the L Η-RH antagonist and its salts. The peptide of the present invention is a homogeneous substance of the luteinizing hormone releasing hormone (LH — RH), which has the following structure P — G1 u — Hi s — T rp_Se r — Ty r — — G1 yL e u-Ar gp r 〇-G 1 y-NH2, [LH-RH, gonadolin]. For more than 20 years, research scientists have begun to seek L Η — R Η decapeptide antagonists with selective potency [M. _Karten and JE Rivier, Endocrine Reviews 1_, 44-6 6 (1 9 8 6)] and so on Antagonists are most interesting for their use in endocrinology, gynecology, contraception and cancer. A large number of compounds have been prepared as potential L H _RH antagonists. At present, the most interesting compounds have been found, which are modified compounds of LH-RH structure. , Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs The first series of effective antagonists were prepared by introducing aromatic amino esters at positions 1, 2, 3 and 6 or 2, 3 and 6. The customary way to write this compound is as follows: show the first amino acid, which replaces the original amino acid in the peptide chain of LH-RH, and the position where the substitution occurs is indicated by a superscript number _. In addition, the expression of ALH — R Η .〃 on the rear represents the LH-RH homogeneous substance that is replaced. Known antagonists are: [Ac_D — Phe (4 — C 1) i.2, D-Tr p3,6] LH-RH (D, Η. Coy et al., Gross, E. a.nd Meienhofer, J . (Eds)

Peptides; Proceedings of the 6th American Peptide This paper size applies the Chinese National Standard (CNS) A4 specification (210X: 297 mm) · '-4-Attachment c: Revised Chinese Specification of Patent Application No. 85114601 September 1998 Presentation V. Description of the invention (51) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs, Table 6: Melting point of the compound according to Examples 5 6 to 8 Example m. P. [° C] Example m, p [° C] Example m, p, [° C] 56 210-213 65 218-222 74 191-193 57 220-223 66 216-219 75 186-188 58 213-215 67 235-238 76 220- 222 59 223-226 68 Gundam 218 77 210-215 60 Gundam 233 69 205-208 78 Gundam 223 6 1 Gundam 237 70 168-170 79 Gundam 226 62 Gundam 221 71 197-202 80 194-197 63 Gundam 220 72 22 BU 226 81 215-222 64 230-236 73 225-228 82 219-222 (Please read the precautions on the back before filling out this page]-丨 Installing _________ 泉 · 丨 This paper size applies Chinese National Standard (CNS) A4 Specifications (210 X 297 mm) -54-A7 B7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs )

Symposium, pp. 775-779 Pierce Chem. Co., Rockville III. (1979): {Ac-Pro ^ D-Phe (4-C 1) 2, D-Na 1 (2) 3, 6] LH-RH (US Patent No. 4,419 '34 7) and [AC — Pro1, D-Phe (4-Cl) 2, D-Trp3 * 6] LH -RH (J. L. Pineda, et al., J. Clin Endocrinol. Meta b. 5 6'420'1983). To increase the water solubility of the antagonist, a basic amino acid such as D-Arg is subsequently introduced in the 6-position. For example [Ac— D — Phe (4 — C 1) 1 2 > D — T r p3> D — A r g6> D — A 1 a10] LH- RH (ORG-30276) (D.H.

Coy, et. Al., Endocrinology 100, 1445, 1 9 8 2); and ...

I 〔Ac— D-Nal (2) 1, D-Phe (4-F) 2, D-Trp3, D — Arg6] LH-RH (ORF 1 8 2 6 0) (]. £ .1? 161 : Et al., 71〇15; 6 ^ 6.11. Nestor, Jr. JJ, Hafez, ESE (Eds). LHRH and its homologs, P p. 1 1-2 2 MTP Prss, Lancaster, UK 1 9 8 4). This equivalent not only has the predicted improved water solubility, but also shows improved antagonistic activity. However, this is equivalent to a strong hydrophilic homogeneous substance with D-A rg 6 and other basic side chains in the 6-position. When administered to rats at a dose of 1.25 or 1.5 mg / kg, it is applied to the face And this paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm). -5-(Please read the precautions on the back before filling the mold page). ~ —ΙΊ .. I ------- ---- Order ---- Printed and repaired on the A 7 消费 年月 日 7 B7 -3-1 5 by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs —) Passive Jane · Explanation Figure 1 shows the competition curve for the binding of cetorelix, compounds of Examples 1, 2, and 56 to human L Η-RH receptor. Figure 2 shows Example 2 The antagonism of the compounds was determined by the concentration of IP 3 in the extract of a human cell line (L3. 5/78) overexpressing the RH receptor. Figure 3 shows the antagonism of the compound of Example 56 as measured by the concentration of IP3 in the extract of a cell line (L3.5 / 78) overexpressing the human LH-RH receptor. Figure 4 shows the inhibition of fluorenone in the blood of rats after injection of the compound of Example 1. Fig. 5 shows the inhibition of fluorenone in the blood of rats after injection of the compound of Example 2. Figure 6 shows the inhibition of fluorenone in blood after rats were injected with the compound of Example 56. The paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page) Ding ______: Yuan loses ________ -'aL2 ^ • 63-f

f US-A A7 B7 V. Description of the invention (3) Temporary edema caused by limbs (F. Schmidt, et al., Contraception 2 9 '2. 8 3, 1 9 8 4: J. E. Morgan, et al. , Contracept i on 29 »283, 1984: JE Morgan, et al., Int. Archs. Allergy. App 1. Immun. 7 0 (1 9 8 6). Other effective LH-RH antagonists are described In WO 92/19651, W Ο 94/193 7 0, WO 92/1702 5, WO 94/14841, WO 94/13313, US-A 5, 3 0 0, 492, 1 4 0, 0 0 9 and EP 0 413 209 Al. The use of some of these antagonists to cause edema effects in rats makes people question their safety when used in humans, which delays the clinical use of these drugs. An antagonist peptide with no side effects is required. According to the present invention, the aforementioned purpose is achieved by a compound of the general formula (I): R2 (Please read the notes on the back before filling this page) K printed by Hansakusha

R1-CO-NH-CH-CO-N |, (Canton 2) n NH R3 (I)

CG -R4 where η is the number 3 or 4, R1 is alkyl, alkoxy, aryl, heteroaryl, aralkyl'heteroaralkyl, aralkoxy or heteroaralkoxy * each may be Unsubstituted or substituted, R 2 or R 3 are individually. Hydrogen atom, fluorenyl group, aromatic compound. The paper size is applicable to Chinese National Standard (CNS) A4 specification (210X 297 mm) A7 ϋ / V. Description of the invention (4 ) Or aralkyl, each may be unsubstituted or substituted, or NR2R3 is an amino acid group, and R4 is a group having the formula (II). O— (CH,) .— Ln — R6 R5 (Η) where ρ is an integer from 1 to 4, R5 is hydrogen or an alkyl group * and R6 is an unsubstituted or substituted aryl or heteroaryl group. Or R 4 is a ring of the general formula (III)

R7 II

X (Please read the notes on the back before filling this page) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (Hi) where q is the number 1 or 2, R7 is a hydrogen atom or an alkyl group, and R 8 is a hydrogen atom or an alkyl group And X is an oxygen or sulfur atom, in which the aromatic or heteroaromatic group may be partially or completely hydrogenated, and the palmar carbon atom may have an R — or S — configuration, and its pharmaceutically acceptable The salt formed by the acid. The preferred combinations of the groups R 1 to R 4 are: a) R1 is benzyloxy, R2 is hydrogen, and R3 is hydrogen, b) R1 is benzyloxy, R2 is hydrogen, and R4 is 4-pyridyl --.------ Order ---------- and c) R2 is hydrogen, R3 is hydrogen, and R4 is 4-fat phenyl This paper applies the Chinese National Standard (CNS) A4 specification ( 210X297 mm) -7-!! Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 ____B7_ V. Description of the Invention (5) Preferred alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl , Isobutyl, third butyl, 2-ethylethyl, dodecyl and hexadecyl. Preferred aryl groups are phenyl, a certain group, phenanthryl and fluorenyl. Preferred heteroaryl groups are pyridyl, pyrimidinyl, imidazolyl, imidazopyridinyl, vermisol, vermizolyl, triazolyl, tetrazolyl, benzimidazolyl, hydroquinone, 2 , 5-dichloropyridine-3-yl and furyl. Preferred hydrogenated heteroaryls are hexahydropyridyl, hexahydropyrazinyl, morpholinyl and pyrrolidinyl. Aryl and heteroaralkyl groups are bonded to the corresponding bonding site through alkylene. Methylene, ethyl, propyl, or butyl are preferred. The preferred substituents are Halogen atoms such as fluorine, chlorine, bromine and iodine, and methyl, ethyl, isopropyl, third butyl, cyano, nitro, carboxylic acid, formamidine, methyl carboxylate, ethyl carboxylate , Ethyl crotonic acid, trifluoromethyl, benzamidine, methoxy, benzyloxy, pyridyloxy, amine, dimethylamino, isopropylamino, fluorenyl and hydroquinone methoxy . Furthermore, according to the present invention, the compound of the general formula (V) is AC-D-N a 1 (2) 1-D (p C 1) Phe2 — D — Pa 1 (3) 3 — Se r 4- T yr 5- D-X xx 6 -L e u7— A r g8— P r o9— D — A 1 a10— NH2 where D — Xxx is the basic paper standard of amino acid of general formula (VI). Applicable to China National Standard (CNS) A4 Specifications (210X297mm). — — Ι! Ίιια) ^. .. / :! (Please read the notes on the back before filling this page) Order ------------ -8- ί ί Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of Invention (6) —HN — CH — CO — 〇—

I (C «2) n 1 i

NH 1. (VI); CO-R4 and η, p, q, R4, R5, R6, R7, R8, and X are as defined above i and the salt formed with a pharmaceutically acceptable acid also achieve the aforementioned purpose. The compounds of the present invention have high antagonistic efficacy and are free of unwanted side effects, especially edema effects. If it does not exist in the form of a salt with a less water-soluble, pharmaceutically acceptable acid, it also has improved water solubility. Furthermore, the compound has a high affinity for the human LH-RH receptor, that is, in Including human mammals, it strongly inhibits the release of gonadotropin from the hypothalamus, inhibits fluorenone for a long time in rats, and minimizes histamine release in vitro. / Preferred compounds of the general formula (I) are: 2 — Ν—Z [£ —Ν — 4 (. 4_fatphenyl) amino-1,4-dioxybutyl] lysamide and 2-Ν —Z [ε— (imidazolidine 2-keto-4 4-yl) methylfluorenyl] lysamidamine ”The preferred peptide according to formula (V) is where Xxx is [eN — — 4 (4- —benzene (Yl) amino-1,4-dioxybutyl] lysylamidinyl or [e-(imidazolidine-2-one-4-yl) methylamidino] lysylamidinyl. The salt formed with a pharmaceutically acceptable acid is preferably less soluble in water. The particularly preferred salt is 4,4 < —methylene-bis (3-hydroxy-2—fruit acid) is also known as the cause Embonic acid or pamoic This paper is sized according to the Chinese National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling " 'This page) Order -9- A7 B7 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of invention (7) acid) β The nomenclature used to define peptides is consistent with the nomenclature described by the IUPAC-IUB Committee (European Journal of Biochemistry 1 9 8 4 * 1 3 8 > 9 — 3 7) > where the same as conventional notation »N-terminal amine group is listed at the left end > and C-terminal carboxy group is listed at the right end 0 L L — R Η Antagonists such as The peptides and peptidomimetics 9 of the present invention include the amino acids that occur in natural and synthetic amino acids »One includes A 1 a > V a 1 9 L e U > I 1 e > Ser» T hr > L y SA rg > AS η »G 1 U» G 1 η) C y S) M ety P he > T yr »P r 0 > T r P and Η i S 〇 each amino acid group The abbreviation of the group is based on the common name of amino acid> is A 1 alanine, Ar S Arginine G 1 y Glycine, Leu Leucine L y S Lysine, P a 9. (3 ) 3 — (3 —pyridyl) alanine Na (2) 3 (2 — a certain group) alanine, P he phenylalanine, (PC) P he 4 — chlorophenylalanine P r 0 proline Acid, Ser serine, T hr threonine% T r P tryptophan and T yr tyrosine. All amino acids described herein are white L — Series Derivatives 9 Unless otherwise stated. For example & gt D — N a 1 (2) is 3 — (2 — a certain group) — D is the abbreviation of alanine> and Ser is L — the abbreviation of serine 0 Other abbreviations used are as follows B 0 C Oxocarbonyl B 0 P _I— fluorophosphate benzotriazole — 1 monooxyl (— · Methylamino) Scale DCC Dicyclohexyl Carbide-Imine (Please read the precautions on the back before filling this page) This paper size applies to China National Standard (CNS) A4 (210X297 mm) -10-A7 B7 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of the invention (8) DC Μ Dichloromethane D d Zn Dimethoxyphenyldimethylmethyleneoxycarbonyl (Dimethoxydimethyl-Z) DIC — Isopropylcarbodiimide DI Ρ Ε AN > N — diisopropylethylamine DMF dimethylformamide F m 0 C fluorenylmethoxycarbonyl Η F liquid ^ frrp m hydrofluoric acid Η 0 Β t 1 —Hydroxybenzotriazole Η PLC high pressure liquid chromatography ΤFA trifluoroacetic acid ζ benzyloxycarbonyl group According to the present invention, the compound of the general formula (I) is obtained by first making two of the three functional groups (α — Amine ε — Amine group and a — carboxylic acid group) with a protective group 9 and then prepared by reacting with the second functional group of Baiyou in an appropriate manner. If appropriate, an intermediate protective group 9 can also be introduced in the first step. Substitution of this with the desired functional group after the second step will yield better results. Appropriate protecting groups and methods for connecting them are known in the art. Examples of protecting groups are described in the principle of peptide synthesis / T Springer Ver lag 19 8 4) »in textbook% solid phase synthesis method f J · M. Stewart and J • D. Υ oung > Pierce Chem. Company, Rockford, III] .984 and G > Barany and R. B • Mer r ifield 'Peptide ff 9th ~~~~ · Chapter 9 Page 1 — 2 8 5, 1 9 7 9, Academic Pre re SSI nc • This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) Please read it first, please note-matters before Fill in and write down: Page 11 Niu C: Revised page of Chinese specification of Patent Application No. 85114601 '1 Supplement I Min_June June V. Description of Invention (9) (Please read the notes on the back before filling this page) The compound of formula (V) can be synthesized by traditional fragmentation or by solid-phase synthesis method according to Merrifield, using D-lysine acid whose side chain has been converted with carboxylic acid of general formula (VII). Sequences are established one by one by D-lysine ° The amidine bond in the side chain causes the decapeptide unit to react with the appropriate residual acid. Thus, according to the present invention, there are three alternative methods for preparing compounds of the general formula (V). The first may include the steps (a) having an appropriate protecting group for the α-amino group and the carboxylic acid group of D-lysine acid or D-ornithine, and (b) the D-lysine acid or D having a protecting group —Ornithine reacts with a carboxylic acid of the general formula (VII) R 4-C Ο Ο Η (VII) wherein R 4 is as defined above, (c) removes the α-phosphonic acid group of the compound obtained in step (b) The protective group is incorporated in position 6 in step (h). It is printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. (D) The D-alanine having a protective group on the amine group is coupled to a resin-type solid. On the support (Merrifield synthesis method), (e) remove the protective group on the amine group of alanine, (f) make the alanine bonded to the solid support and the proline acid having a protective group on the nitrogen atom Reaction, (g) removing the protecting group on the nitrogen atom of proline,

(h) Use amino acids 1 to 8 of formula (V) in the order of 8 to 1; use the modified D for position 6 described in step (c). D This paper size applies Chinese National Standard (CNS ) A4 specification (210 x 297 mm) -12-A7 B7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (ίο) Monolysine or 〇_Ornithine, repeat steps f) and g) (I) Remove the compound obtained in step (h) from the support and purify it if necessary (e.g. HPLC), (j) if necessary, react with a pharmaceutically acceptable acid, the acid is Acid is preferred. According to the second generation, the method for preparing the compound of general formula (V) includes the following steps: (a) coupling D-alanine having a protective group on the amine group to a carrier suitable for solid phase synthesis, (b) removing the protective group located on the amino group of alanine, (c) reacting the alanine bonded to the resin with proline acid having a protective group on the nitrogen atom,., (d) removing the (E) using amino acids 1 to 8 of the general formula (V), repeating steps (c) and (d) according to the order from 8 to 1, (f) self-supporting The compound obtained in step (e) is removed, (g) is reacted with a carboxylic acid of formula (VI I) R4-COOH (VII) where R 4 is as defined above, and (h) if necessary, it is compatible with a pharmaceutically acceptable Alkali reaction, the acid applies the Chinese National Standard (CNS) A4 specification (210X297 mm) at this paper scale '_ (Please read the precautions on the back before filling, this page) Order «dm vmnQ .IBK m ^ i ^ i—- As Α7 δδ. 6. 8 Β7_ 5. Description of the invention (11) Inbornyl acid is preferred. The third variation of the production method of the compound of general formula (V) includes the following steps (please read the precautions on the back before filling this page) Step (a) coupling D-alanine with a protective group on the amino acid On a support suitable for solid-phase synthesis, (b) remove the protective group on the amine group of alanine, (c) make alanine bonded to the resin and proamine having a protective group on the nitrogen atom Acid reaction, (d) removing the protecting group on the nitrogen atom of proline, (e) using amino acids 6 to 8 of the general formula (V), and repeating steps c) and d in the order of 8 to 6 ), (F) removing the e-amino protecting group at position 6 from D-lysine or D-ornithine, and reacting with a carboxylic acid of formula (VI I), R 4-C 〇 Ο VII (VII) Among them, R 4 is as defined above. The consumer cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs printed (g) to remove the protective group on the α-amine group of D-lysine or D-ornithine. (H) Use the general formula ( V) For amino acids 1 to 5, repeat steps c) and d) in the order of 5 to 1. (i) Remove the compound obtained in step (h) from the resin, and purify the other (e.g., PLC), (j) If necessary, react with a pharmaceutically acceptable acid, which is suitable for China at the paper scale National Standard (CNS) A4 specification (210 X 297 mm) -14-A7 B7 V. Description of the invention (12) Inbernamic acid is preferred. (Please read the notes on the back before filling out this page) The preferred formula (VI I) is carboxylic acid imidazolidine 2-ketone-4 monocarboxylic acid and N- (4-phenylphenyl) amino-4 Oxybutyric acid. Compounds of formula (V) are synthesized according to known methods, such as, for example, pure solid phase technology, partial solid phase technology, or traditional solution coupling methods (see M. Bodanszky, ^ Peptide Synthesis Principles〃 JM Stewart and JD Young, Pierce Chem Company, Rockford, III s 1 9 8 4 and G. Barany and RB Merrifield 'Peptides' Chapter 1 'pp. 285, 1979, Academic Press Inco Traditional solution synthesis is detailed in " ^ Methoden der' Organise hen Chemie [Organic. Chemical Method] (Houben-Weyl), Syuthe.se von Peptiden [winning synthesis method] + E. Wlinsch (edition) 1 9 7 4.. G e 〇rg T hieme V er 1 ag , S tu 11 gart, FRG. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, a step-by-step synthesis method, for example, first covalently bonding the carboxy-terminal amino acid (the α-amine group of which is protected) to those commonly used in such cases On the insoluble support, remove the α-amino protecting group of this amino acid, so that the next protected amino acid is bonded to the resulting free amino group via its carboxyl group. According to this method, follow the correct sequence, Step by step For other amino acids of the peptide, after binding all the amino acids, the final treated peptide is removed from the support, and other lateral functional group protecting groups that are present are removed as appropriate. The stepwise reduction method is customary. The method starts from the synthesis of an appropriate amino acid protected according to the customary method. Similarly, when using an automatic peptide synthesizer, such as Labortec S P 6 50 of Bachem, Switzerland, a commercially available protected amino group can be used. This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) -15-Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 ___ B7 ^ ____ V. Description of the invention (13) Inter-bonds of individual amino acids The binding is carried out according to the method used in this case, and the following are particularly applicable:. The method of symmetric anhydride synthesis in the presence of dicyclohexylcarbodiimide or diisopropylcarbodiimide (DCC'DIC). Imine method. Carbodiimide-hydroxybenzotriwa method (Reference, Peptide, Book 2, Edited by E. Gross and J. Meienhofer). For the binding of arginine, carbodiimide is used The law is better. For the amino acid, symmetrical or mixed anhydride method is usually used. In fragment coupling, use acid coupling method (performed without racemization) or DCC — 1 -hydroxybenzotriazole or DCC — 3 — The hydroxy-4-oxy-3,4-dihydro-1,2,3-benzobenzozine method is preferred. Fragile active esters can also be used. In terms of the stepwise condensation of amino acids, particularly suitable active esters are N = protected amino acids such as, for example, N-hydroxysuccinimide or 2,4,5-trichlorophenyl ester. Ammonialation can be catalyzed very quickly by N-hydroxy compounds, which have about the acidity of acetic acid, such as, for example, 1-hydroxybenzotriazole. Existing intermediate amine protecting groups are groups that can be removed by dehydrogenation, such as, for example, benzyloxycarbonyl (= Z group) or groups that can be removed by weak acids. Examples of protective groups used for α-amino groups are: tertiary butoxycarbonyl, benzyloxycarbonyl, or sulfanylthiocarbonyl (with p-bromo or p-nitrobenzyl as appropriate), trifluoroacetamidine, benzene Dimethylfluorenyl, o-nitrophenoxyethylfluorenyl, trityl, p-toluenesulfonyl, benzyl, and benzene rings. Applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) for this paper size (please first Notes on the back of the reading (ar page) nn 1— tm — ^ i ^ i ^^^^ 1 > ι ^ ϋΒ I --- Jtm * mm ·-: i'4 i -16-A7 B7 V. Description of the invention (14) Substituted (p-bromo or p-nitrophenyl) benzyl and α-phenethyl. Reference is also made here to the book by Jesse P. Greenstein and Mil.ton Winitz, Amino Acid Chemistry, New York 1 961, John Wiley and Sons, Inc., Book II, e.g., pages 8 8 and below, and peptides , Book II, Ed. E. Gross and J. Me i enhofer, Academic Press, New York. Protecting groups such as ft are also basically suitable for protecting other functional side groups of corresponding amino acids (0 Η group, N Η 2 group). The hydroxyl group (serine, threonine) contained is preferably protected by benzyl and similar groups. Other amine groups not located at the α-position (for example, amine groups at the omega-position >> guanidine of arginine) are preferably protected orthogonally. The reaction used for amino acid bonding is performed in a conventional solvent or a suspending agent (such as dichloromethane). If necessary, dimethylformamide can be added to improve the solubility. As far as the 114-C0 group is introduced by reacting an amine group of lysine with a carboxylic acid of the general formula (VII), the same method as described above is basically suitable for bonding an amine. However, the condensation characteristics of carbodiimide are used, such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide and 1-hydroxy-monobenzotriol. A suitable synthetic support is an insoluble polymer, such as a polystyrene resin in the form of beads, which can swell in organic solvents (such as a copolymer of polystyrene and 1% divinylbenzene) in methyldibenzene Protected decapeptide ammonium is synthesized from methyl ammonium resin (MB ΗΑ resin ', which is a polystyrene resin with methylbenzyl amidoamine group). After excision of the support with HF, the peptide is expected to be produced. ， C-terminal pinamine functional group, this synthesis method can be used _ This paper size applies Chinese National Standard (CNS) A4 specification (210X297 public director) Please read the note at the back

I i Printed by the Employees 'Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs -17-A7 B7 V. Description of the Invention (15) The following flow chart is performed: Printed by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economics (Please read the precautions on the back before filling out this page)

This paper size applies to China National Standard (CNS) A4 (210X297 mm). 1 18 —

A B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (丨 c) Flow chart of peptide synthesis method. Functional solvent / reagent (v / v). Time 1 Wash methanol 2 X 2 minutes 2 Wash DCM 3 X 3 Minute 3 Remove DCM / TFAC1: 1) 1 X 30 minutes 4 Wash isopropanol 2X 2 minutes 5 Wash methanol 2X 2 minutes 6 Wash DCM 2 X 3 minutes 7 Neutralize DCM / DIPEAC 9: 1) 3 X 5 minutes 8 Wash methanol 2X 2 minutes 9 Wash DCM 3X 3 minutes 10 STOP Add Boc-As 11 coupling in DCM + DIC + HOBt-about 90 minutes 12 Wash methanol 3 X 2 minutes 13 Wash DCM 2 X 3 minutes (Please read the back Note: Please fill in this page again.) This paper size is applicable to China National Standards (CNS) A4 specification (210 X 297 mm)-1 5-Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Invention Description (17) Να —B oc protected amino acid in CH2Ce2 / DMF in the presence of diisopropylcarbodiimide (DIC) and 1-hydroxybenzotriazole (HOB t) in 90 minutes Coupling with triple Molar excess in time, by CH2C 2 of 50% trifluoroacetic acid (TFA) acting over a half hours, the protecting group is removed B0C. To confirm complete conversion, the Chloranil test according to Christensen and the Kaiser's ninhydrin test can be used. The free amine function is based on the tritiation of CH2C2 with a five-fold excess of acetamidoimidazole. The sequence of the peptide synthesis reaction on the resin follows this flow procedure. When the peptide bonded to the resin is removed, each final product of the solid phase synthesis is dried in a vacuum at P 2 0, and a 500-fold excess of HF / anisole 10: 1 (v: v ) For 0 minutes. After distilling off HF and anisole in a vacuum, a white solid was obtained by stirring with anhydrous ether; the polymer support was removed by washing with a 50% strength acetic acid aqueous solution. get on. The acetic acid solution was carefully concentrated in vacuo to obtain individual peptides in the form of a highly viscous oil, which was converted to a white solid under cooling after the addition of anhydrous ether. Further purification was performed by conventional methods of preparative high pressure liquid chromatography (HPLC). The conversion of the peptide into its acid addition salt proceeds by reacting it with the acid in a known manner. Conversely, free peptides can be prepared by reacting their acid addition salts with bases. The peptide inbornyl salt can be prepared by reacting the trifluoroacetate (TFA salt) of the peptide with free inbornyl acid (pamoic acid) or the corresponding disodium inbornyl acid. In order to complete this reaction, the peptide TF Α salt is in an aqueous solution, and the Chinese National Standard (CNS) A4 specification (210X297 mm) is applied to the paper size of paper. — ~ -20-(Please read the precautions on the back before filling in { (This page)

A7 B7 V. Description of the invention (18) The solution of disodium nitrate in a polar aprotic medium (preferably dimethylacetamide) is treated, and the pale yellow precipitate formed is isolated. The following examples are intended to illustrate the invention, but not to limit it. Please read the notes and events on the back before implementing the examples

Ac — D -N a 1 (2) -D (PC 1) P he — D ~ P a 1 (3)-Ser — T yr-D — [e -N ^-(imidazolidine-2 monoone- 4 1 base) Formamyl] — L ya — Leu —

Ar g — P r 〇 — D — A 1 a — NH The consumer cooperation of the Central Bureau of Light Industry of the Ministry of Light Industry is printed according to the flow chart * at 5 1. 08mmo 芡 / g) ~~ D — L ys ( B oc) — 0 after the Boc group, then triplicated with 3 times percarboxylic acid. After using 20% hexahydropyridyl, according to the flowchart, after drawing the body at N, 5.2 g of crude peptide was obtained and purified. After subsequent freeze-drying, C74H97Ni8015Cj? HP's FAB-MS 15 14 (1512. 7) and the corresponding 1H-BHA resin (filling density were synthesized. Lysine was coupled in the form of Fm 0c Η and removed in the side chain The amount of imidazolidine-2-ketone-4 dipyridine / DMF was used to remove the Fmo c end-cap for chain extension. The polymer was removed, and 2.1 g of the experimental LC homogeneous product was obtained by the HPLC standard, which had the correct M + Η +) (Calculated value-NMR spectrum. 1H-NMR (500MHz, DMSO-de, 5 expressed in PP m): Order '4! This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm)

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs I A7 B7 V. Description of Invention (19) 8.56, m, 2H, aromatics. H; 8. 08, m, 1H, aromatics. H; 7 · 81, m, 1H, aromatics. H; 7.73m, 2H, aromatics. H; 7.66, m, lH, aromatics. H; 7.60, s, lH, aromatics. H; 7.44, m, 2H, aromatics. H; 7.30, d, lH, aromatics.矸; 7.25, and 7.18, 2 (1,2 father 211, aromatics. H p-C 1-P he; 6.97 and 6.60, 2d, 2x2H, aromatics. Η T yr; 9.2- 6.3, several signals, ammonium NH; 4.8 — 4.0,-several m, C α — Η and aliphatic. Η; 2. 1 — 1. 1, several m, residual aliphatic. Η; 1 70, s, 3H, ethenyl; 1.22, d, 3H, C; 5-H Ala; 0.85, dd, 6H, C5-H Leu Example 2

Ac-D-Na 1 (2)-D (PcL) Phe-D —Pal (3) — Ser— Tyr— D [e— N1 — 4 — (4 —fluorenyl) amino — 1, 4 — di Oxybutyl] —Lys — This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) (Please read the precautions on the back before filling out this page), JI I- nn I— iHS 、 一 IN I! --- 1 n ^ in -22-V. Description of the invention (20) A7 B7 L e UA r SP Γ 0 DA 1 a-N Η 2 0, 7 mm 0 9. (1 • 0 3 G) + A c —D — N a J2 — D — (Ρ C 9.) ρ he — D — Pa 1 —Ser — T yr — D — L y S — Le U — A rg-ρ r 〇— D — A 1 a — NH 2 and] L _ 0 丨 m m. 〇1 (0 · 2 7 g) (4 -aminophenyl) amino — 4 -oxybutyric acid at 1 • 〇mm 0 Si (0 • 1 6 g) 1 —Ethyl — 3 — (3 —Dimethylaminopropyl) carbodiimide and 1 • 0 mm 0 9. (0 1 6 s) 1 — Hydroxybenzotriazole is present 'The reaction was carried out in the initial distillation of DM. After 24 hours in a vacuum, the residue obtained by removing the solvent was dissolved in water, and the solution was freeze-dried. The crude reaction product (1 * 6 3 g) obtained by the preparation was purified by reverse phase Η PLC to obtain a total of 0 • 6 1 g experimental formula C 8 1 Η 10 4ν α 9 〇 1 5 c 9. Η [PLC homogeneous product, which has positive-correct FA Β-Μ S * 1 6 1 8 • 7 (M + Η + ) (Calculated value 1 6 1 7. 7) and corresponding. And 1 Ε [— NMR spectrum < > (Please read the notes on the back before filling in this page) Order the stamp of the staff consumer cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 6 d Iο SΜ D z ΗΜ ο ο 5 / IN RΜ Ν I Η

s S ο ο, '.物 物 5 Η 物 族 Family Group 1 Ν 芳芳. Aromatic ,, 9 amines and fluorene and benzene, 11 fluorenyl hydrazone ,, 1 脒 2mm • ♦ t I 9 f,) s 4 moo shown,, 2 8 Table 4H0.. M · X 6 8 7, ·· t · I PI s 8 Η H 8 8 and Η 2 and A s N c

I centimeter 7 9 2 X A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (21) 1 Η 1 7 7 3 > m) Aromatics o 1 1 1 Η 1 7 • 6 1 9 S 9 1 Η »Aromatics 〇1 1 Η» 7 4 4 9 m 9 2 Η j Aromatics 0 II Please 1 Η 7.. 3 0 > d 9 1 Η Aromatics 0 News I Read | Η t 7. 2 5 and 7-2 0 2 d 4 Η 9 Aromatics 〇 1 I 1 Η (Ｃ C 1) P he «Note # 1 7 0 and 6 6 2 D > 4 Η Aromatics The term i Η Τ yr writes: this page 1 8 3 a 7 • 2 several letters Wu nr < fi · amine-N Η S_✓ 1 4 7 3 — 4 • 2 9 several multiple lines 1 1 C a — Η »1 1 4 1 3 9 m 9 1 H 9 C a — Η > Order | A 1 a 1 I 3 • 7 8 — 2 4 Several multiple lines >, ^ 1 1 IC β — Η and aliphatic 0 1 1 '1 Η t 1 • 7 2 »s 3 Η s acetamyl * S 1 • 2 2 td, 3 HC β A 1 af 1 0 • 8 5 9 dd > 6 Η C δ L eu 1 Example 3! I 0 • 5 g (0 3 mm 0 9.) The peptide L of Example 1 Η-1 1 | R Η Antagonist solution In 50 m H 2 〇 'by reacting with 0 • 1 3 0 g (0 3 mm 0) disodium pamoate in 2 mj? Aqueous solution 1 1, 1 1 was converted into the peptide inbornyl acid The salt is rapidly formed in the form of a yellow precipitate from 1 1 this paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) -24-2 8 1 g fine crystalline yellow-green powder, Imborni A7 B7 5 2. Description of the invention (22) The solution precipitates. 0% acid content 3 3% »EMM 4 0.3g (0. 17mmoj2) The peptide LH-RH antagonist of Example 2 was dissolved in 5miT dimethylacetamide, and 0.195g by 2mj? Aqueous solution. (0.45mmoj?) The disodium pamoate reacted to convert the peptide inbornyl salt, and a yellow precipitate obtained after adding 50 m of sense H2O. 0.30 g of a finely crystalline yellow product was obtained with an inulinic acid content of 20%. The compound of the general formula I can be obtained according to the following schematic diagrams 1, 3, 4 and 5, and the three functional groups R1, R3 and R4 are changed integrally. Schematic 1 shows the synthesis method of the compound of Example 1: Schematic diagram 1 Please read the back. · Note for refilling, write: This page is printed on the paper printed by the Central Consumers Bureau of the Ministry of Economic Affairs. ) Α4 specifications (210 × 297 mm) -25-A7 B7 V. Description of the invention (23) Ου

HOOC

Ου

V

1) Benzotriazole-oxyloxy-tris (dimethylamino) iron (BOP) hexafluorophosphate 2) N-methylmorpholine 3) 2n NaOH 4) CF3C00H Please read the back first. Note the things to write again 'This page

NH, printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs. 0 _ # · _ [!} 1 The general method for preparing compounds of general formula I was used as the basis of general formula I and the schematic diagram 1 and replaced by group 1 ^ 4. The carboxylic acid R. 4 _ C ο ο Η, if R 4 is a basic group, it can also exist in the form of a salt, such as the hydrochloride, bisulfate or acetate, under the exclusion of moisture and stirring, dissolved or Suspended in non-polar or dipolar aprotic organic solvents such as, for example, tetrahydrofuran, dioxane, methyl tertiary butyl ether, toluene, dimethylformamide, dimethylacetamide, N-methylpyrroline Ketone, dimethyl sulfene or dichloromethane, with alkali treatment as an acid absorbent under stirring, such as, for example, using diisopropylamine, triethylamine, N-methylmorpholine, two paper standards applicable to China National Standards (CNS) M specifications (210X297 mm) -26-A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (24.) Methylpyridine or pyridine. A mixture of Z- (L) -lysamidamine in diduent is then added, and the appropriate duty is the aforementioned one for dissolving the carboxylic acid R4-C ◦ 0H substituted with the group R4. The pH of the reaction mixture is adjusted to pH 6.5 to 9.0, 7.0 to 8.5, and particularly preferably 7.0 to 7.5. Finally, with further stirring, a solution of a coupling reagent is added to the reaction mixture, such as benzotriazole hexafluorophosphate-1 -yloxy-tris (dimethylamino) iron (BOP), or six The benzotriazole fluorotrifluoro-l-yloxy-tripyrrolidinyl scale (Py BOP) or dicyclohexylcarbodiimide (DCC) is adjusted to the aforementioned pH range again after a short time. The suspension is stirred at, for example, 0 to 80 ° C for 1 to 15 hours, preferably 10 to 50 ° C, more preferably 20 to 30 ° C, and then filtered by suction, washing the solid, and concentrating the filtrate in vacuo to dry. The residue is crystallized by rubbing with an organic solvent, such as toluene, tetrahydrofuran-acetone, methyl ethyl ketone, or isopropanol, or by recrystallization, distillation, or flash chromatography on silica gel or alumina column chromatography. . The eluent used is, for example, a mixture of dichloromethane, methanol, ammonia (25%) at a ratio of 85: 15: 1 (volume / volume), or dichloromethane, methanol, ammonia (25%) at 80:25 • Mixtures at a ratio of 5 (volume / volume). Trifluoroacetate synthesis method: The compound purified according to the aforementioned method is dissolved in a protic or aprotic solvent, for example in an alcohol such as methanol, EtOH, isopropanol, or in a cyclic ether such as For example, tetrahydrofuran or dioxane, and use 2 N hydrogen. The paper size is applicable to Chinese National Standard (CNS) A4 specification (210X297 mm) " " (Please read the precautions on the back before filling this page) J. Order- -Λ Printed by the Consumer Cooperative of the Central Bureau of the Ministry of Economic Affairs of the Consumer Cooperative A7 B7 V. Description of the invention (25) The sodium oxide solution is adjusted to pH 10-11. The precipitated solid was filtered off by suction, washed, dried in a vacuum, and used at a temperature of 10-80 ° C (preferably at 20-400 ° C) in an ethanol solution using an equal amount or 2 -4 times molar excess of trifluoroacetic acid. The solution was at 0-4. (: After standing for 24 hours, the desired trifluoroacetate is precipitated, filtered by suction, and dried in the air. According to this basic general method as a synthetic schematic diagram 1, according to the description of Example 5 below and below Table 1 Synthetic compounds: Example Fi 2 ~ N — [fluorenyl ferroyl] —g—N—Γ 5 ~ [(4-fl and phenyl_.yl) amino] —5_oxypentyl] — L, _Lysamine triamine acetate 5g (17.5mmoj?) 5 — [[4-mono (aminoiminomethyl) phenyl] monoamino] -5 -oxyvalerate Under stirring and dehydration, suspend in 20 Omj? Dimethylformamide, and treat with 3. 85mJ? (35. Ommoj?) N-methylmorpholine. Add 5. 5 3g (17.5mmo 芡) A mixture of Z- (L) -lysamidamine hydrochloride in 10 Omj? Dimethylformamide, using N-methylmorpholine to adjust the pH to 7.0-7.5. Finally, add 9. 73g (21.9mmoj?) Hexafluorophosphate benzotriazole_1-ylmonooxytris (dimethylamino) scale solution, after 10-15 minutes, adjust the pH to 7.0 -7 · 5. Stir the yellow suspension at room temperature under continuous pH detection, The pH should be 7.0-7.5. After this paper size, the Chinese National Standard (CNS) A4 specification (210X297 mm) applies. 28_ (Please read the precautions on the back before filling this page) Set the central standard of the Ministry of Economic Affairs A7 printed by the Bureau-Industrial and Consumer Cooperatives _____B7__ V. Description of the invention (26) 3-4 hours, the colorless precipitate was filtered off by suction, washed with dimethylformamide twice, and the yellow filtrate was evaporated to dryness. After a total of 5 x 4 0 m and methyl ethyl ketone extraction, after every five solvent treatments, the methyl ethyl ketone phase was decanted and discarded. The remaining crude product in the crystalline form was filtered by suction and filtered at 30 mi Methyl ethyl ketone was washed and dried at room temperature in a vacuum. The solid was dissolved in about 50mj? Ethanol, adjusted to pH 10-1 with 2N sodium hydroxide solution. The precipitated alkali was filtered off by suction, water and ethanol Wash and dry in vacuum at 35 ° C. Yield: 5.5 g (62% of theory) = 帛 7, ._ Test: 5.5 g of alkali in ethanol suspension at 60 ° C , Treated with 5 times the molar amount of trigas acetic acid. The solution was stored at 4 ° C overnight, and the trifluoroacetic acid trifluoride was filtered by suction at 35 ° C. Dried in vacuo ^ 'Yield: 5.9g (87.7% of theory)

Melting point: 1 8 5 ° C Elemental analysis: Calculated value C 5 3 · 8 4 Η 5.65 Ν 13.45 Experimental value C 54.11 Η 5.74 'Ν 13.33 Spectral (50 0Mhg DMSO — dR, (5 Applicable on this paper scale? (CNS ) A4 · (210x297 public thin) _ 9q _ '(Please read the precautions on the back before filling this page) \ 丨 \ · 1 -.- 1 1-· -s • · 1 -n. Ν .Ί " • re. A7 B7 V. Description of the invention (27) ppm means: 10. 47, s, 1H, panitaniline, 9. 14 and 8. 8 2s, NH #, 7.82, m, lH, lys-e — NH , 7. 79 and 7. 46, 2s, aromatic. Η, 7. 27 and 6.93 2s, 2H, CON2, 7.20, d, 1H, urea NH, 5.0, s »2H, benzyl H, 3.89, m, lH, Ca-H, 3.0 and 2. 58 and 2. 40, 3 m, all 6H, aliphatic. 1 ^, 1.60 — 1.20, 4111, all 611,. 犷 6111. aliphatic. Η R2 R1-CO-ΝΗ -CH-CO-Ν R3 (广 2) n

NH

I CO-R4 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page) C Formula I) According to the aforementioned method, prepare the other compounds listed in Table I below, where n is equal to 4 . Table 1: According to Synthetic Schematic Diagram 1 and according to the general formula I-2, e—N—substituted L—lysamidamine derivatives (η in all examples are equal to 4) This paper size applies to China National Standard (CNS) A4 specifications (210 X 297 mm) -30-A7 B7 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of the invention (28) Example le R'-CO R2 / R3 R4

—II In -....... ^^ 1-.-11 —1-i ,, ~ —II —: m (Please read the notes on the back before filling this page) Order ---- ---- «m IB > ^ —mu · This paper size is applicable to China National Standard (CNS) A4 specification (210X297 mm) -31-

32 A7 ___B7 V. Description of the invention (30) 袠 丄 (continued) 丨 Printed and implemented by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs J le R1-CO R2 / R3 R " 25 H / H 0 \-二 / \ _ / 26 27 righteousness. 28 ο C, CH: again Np cooet 29 Λι less NH,%, NH 30 'Co CoCHJJJ 31 iy WN 32 name 〇_ / 飞, \ _ / H \ = / 33 "' 人 " ~ 〇 ~ < ^ 34 'National Standard (tNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling out this page) Order — -33-A7 B7 V. Description of the invention (31) Melting points of the aforementioned example compounds See Table 2 below. Table 2: Melting points of the compounds of Examples 5 to 34 The paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) -34-A7 B7 V. Description of the invention (3〇 Economy Example mp [° C] Example mp [° C] Example mp [° C] 5 185 15 225 25 Pasty residue 6 185 16 211-214 26 205-210 7 216-220 17 183-186 27 172-177 8 225 18 (oil) 28 227-230 9 217-220 19 slurry residue 29. 225-229 10 218-222 20 (oil) 30 233-235 11 208- 212 21 (oil) 31 215-218 12 (oil) 22 (oil) 32 155 13 232-236 23 Paste residue 33 (oil) 14 194-198 24 (oil) 34 (oil) ( Please read the notes on the back before filling this page.) Thread #. This paper size is applicable to China National Standard (CNS) A4 (210X297mm) -2 »5- A7 B7 V. Description of the invention (33) The precursors used to formulate the compounds of the general formula I are shown in Table 1. As shown in Table 1, the starting compounds Z — (L)-acid amines at the final stage of the synthesis of Examples 15 to 34 are commercially available. For other starting materials, the substituted, aryl, or aryl aryl alkanoates from the synthetic scheme 1 can be prepared by known methods in the literature as in the synthetic scheme 2 (p. R. Bovy , Organ. Che m. 5_§_, 7 948 (1 993)). Schematic 2 Please 7 ^, read the notes of the back and fill in, write this page 〇 person α-νη2 + (CH2) p ο ο

(CH2) ^ NH

ο HOOC- ^ Order A = aryl heteroaryl ρ = 2-5 A = aryl, heteroaryl p = 2 — 5 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs for the synthesis of aromatic or Heteroaromatic amines A — NH 2 are commercially available; Example 28 Aminoimidazo [1,2-a] D ratios based on 8 compounds can be synthesized according to methods known in the literature (r. Westwood, J. Med. Chem 31-1098 (1988)). Aaryl 'mono or ~ heteroarylaminooxyalkanoic acid, which has been previously designed as a precursor, may be additionally prepared by monomethyl alkane dicarboxylate (eg suberic acid mono Methyl ester and adipic acid monomethyl ester), with aromatic or heteroaromatic amines in boiling alcohol (such as boiling ethanol or butanol), or optionally in aromatic solvents (such as the paper size of this example) Applicable to China National Standard (CNS) A4 specification (210X297 mm) -36-Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (34) In toluene or xylene) The case is prepared in an autoclave at the boiling point of the solvent using a pressure of up to 5 Ob ar, prepared by the decomposition reaction of the amine group, in a vacuum The reaction solution was concentrated in water, and the residue was purified by crystallization of methanol or ethanol or by column chromatography. The eluents used were, for example, dichloromethane, methanol, gas (25%) ratio 85: 15: 1 (volume / volume) ), Or a mixture of dichloromethane, methanol, and ammonia (25%) in a ratio of 80: 25: 5 (volume / volume). An alternative method for preparing compounds of general formula (I), where R1 is fluorenyloxycarbonyl, R 2 and R 3 are hydrogen atoms, as follows: 1. α-carboxylic acid group is aminated with amidine. 2. · ε-amino group is protected by ζ group. 3. α-amino group is protected by B 0c group, This results in the slower selectivity of the amine protecting group. / 4. Removal of the Z group located on the ε ~ amine group. 5 · Leads the desired group R4-CO-to the ε -amine group. 6. Removal of the Boc group located on the amine group. 7. The group provided on the α-amine group. Other compounds of the general formula I can be obtained according to the schematic diagram 3 below: Synthesis method of the compound representing Example 35: This paper Standards apply to China National Standards (CNS) Α4 specifications (2 丨 0'χ297 mm) H,: 1-1-»^^ 1 ill I. .Ϊ IV I--II--In--aJ--i . ^ 1 1 - - I - (Please read the back of the House precautions to fill out this page) # · -37 - A7 B7 Ministry of Economic Affairs Bureau of Standards employees consumer cooperatives printed five or description of the invention (35) Figure 3 shows actually

Please read the precautions on the back first, and then go to 'Mai' on this page. The paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) 38-A7 B7 V. Description of the invention (36) Prepared according to the diagram 3 General Method for Compound I of Formula I Phase 1 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) at a temperature in the range of 30 ° C to 30 ° C Down (between -20 ° C and 20 ° C is better, between -15 ° C and 5 ° C is particularly good), Z — Lys (Boc) — 0H and alkali, Examples include triethylamine, diisopropylamine, N-methylmorpholine, N-ethylhexahydropyridine, and aliphatic or aromatic carbonyl chlorides, such as acetamidine chloride, isobutylammonium chloride, isopentamidine chloride, trimethyl Acetyl chloride, pyrene chloride or 4-methoxychloro chloride, added to dipolar aprotic or non-polar organic solvents, such as, for example, tetrahydrofuran, dimethylarsox, dimethylformamide, acetonitrile, acetic acid Ethyl acetate, dimethylacetamide, N-methylpyrrolidone, dioxane, toluene, ether, dichloromethane or chloroform ^ After a certain period After a period of time, for example, 30 minutes to 3 hours, add amine to a dipolar aprotic or non-polar organic solvent (such as tetrahydrofuran, dimethyl asus, dimethylformamide, acetonitrile, and ethyl acetate under vigorous stirring). , Dimethylacetamide, N-methylpyrrolidone, dihumane, toluene, ether, dichloromethane or chloro (form). The suspension is at a temperature between 30 ° ( : Stir to a temperature in the range of 30 ° C (preferably between 20 ° C and 20 ° C, especially between 15 ° C and 5 ° C) and stir for 1 to 2 hours. After the reaction, the base in the form of hydrochloride is filtered off by suction and the solvent is concentrated. The oily residue is aprotic or non-polar organic solvents (such as diethyl ether, diisopropyl ether, methyl tertiary butyl ether, petroleum ether, Toluene, xylene, pentane, hexane) treatment. The solution is stirred for a normal time, such as 30 minutes to 3 hours, until a white powder is precipitated. The precipitate is filtered off by suction and dried. This paper size applies to Chinese national standards (CNS ) A4 specification (210X297mm) -39-A7. B7 V. Description of invention (37) In the second stage of the Ministry of Economic Affairs The Z-Lys (Bo c) amine obtained according to the method of the first stage described above was printed by the Central Consumers ’Cooperative Consumer Cooperative at a temperature between 20 ° C and 30 ° C (between 10 ° c and 20 ° C, preferably between -5 ° C and 5 ° C) Dissolved in trifluoroacetic acid, stirred for 15 minutes to 1 hour. Concentrated excess trifluoroacetic acid, oily residue The substance is treated with a dipolar aprotic or non-polar organic solvent, such as, for example, dimethylformamide, dichloromethane, tetrahydrofuran, acetonitrile, N-methylpyrrolidone, ethyl acetate. Desired acids, bases (such as, for example, diisopropylethylamine, N-methylmorpholine) and appropriate coupling agents (such as, for example, B Op) are then added to the dipolar aprotic or non-polar organic solvents. , PyBOP, DCC), and the solvent is, for example, dimethylformamide, dichloromethane, tetrahydrofuran, acetonitrile, N-methylpyrrolidone 'ethyl acetate. The reaction is carried out at a temperature from −101 to 100 ° C, with 0. (: It is preferably to 80 ° C, especially between 10 ° C and 35 ° C. After a reaction time of 1 to 5 hours, the solvent is concentrated after being left at room temperature for 24 hours. Use Organic solvent precipitates the residue 'The solvent is, for example, water, isopropanol, dichloromethane or diethyl ether. The crude product is purified by chromatography on a silica gel column. According to the basis of the synthetic scheme 3, steps 1 and 2 are used. General method: Synthesize the compounds in Table 3 below, η are all equal to 4. This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) -40-A7 B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

(Please read the precautions on the reverse side before filling / this page) Binding order specification Α4 \) / s N c quasi-standard Tsukakuni country r; fl X X ο c >-~ Public A7 B7 V. Description of the invention (39 ) Table __a_ (continued) »mj R1-CO R2 ___ 9 3

H

40 1 4

____ \) / c _I ^ DI > n ^ — HB (—E ^ i. am tr-f · 丨 · IB Mfl ^ n M,.:., 4 (Please read the precautions on the back before filling in ^ this page ) 42 43

-Order --- 5 4 Central China Bureau of Standards, Ministry of Economic Affairs, Consumer Cooperatives, China 46 47 8 4 Appropriate Standards, National No.1 High School, Moderate Rule-Paper

S Ν C Μ

c 0

X ° | centimeter 2 4 A7 B7 V. Description of the invention (4〇) Table_2_ (continued)

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (53 cases) ZE- Ν- α- Ν 1 Amine Segment Stage 1 First Base Gas ε

N 4

4 / (V-based phenylfluorenylamino amines

N-based methylpyridine hydrazone I 3 This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) -43-2 A7 B7 V. Description of the invention (41)

N L v s (B 〇 c)

N (3-Pyridinyl) pyrimidine (α-N-Z_ [g-N -Third butanecarbonyl 1 lysine_ (IV -Pyridinylmethyl)) pyrimidine 15 ° (: 42 (1〇 111111〇 and) commercially available 2-Ly and 1 m 芡 1 .m and -1 hydrochloride dried s (B oc)-0.26 g (10 m in tetrahydrofuran. After 0 8 g ( 10mm tetrahydrofuran was stirred for 1 to 2 at 5 ° C before cooling, and then the tetrahydrofuride was evaporated. The solution was stirred until H, lg (lOmmoj?) Triethylamine m 〇 $) trimethylacetamidine was added to 6 0 3 After 0 minutes, a solution of 3- (aminemethyl) pyridine at 20 to 10 ° C was added with vigorous stirring. The suspension was allowed to stand for hours. The triethylamine salt was filtered off under suction at low temperature. The oily residue is a white powder with 10 Omj? Diethyl. The precipitate is drawn out by the suction furnace, and please read the precautions before filling in / this page. The output printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economy: 4g (Theoretical value is 85%). Stage ϋ— ~ (α ~ N — Z “e— N — 4 — (4-benzylbenzene) —amine oxybutyrate] lysine mono-N— (3-pyridine A) Amine 2g (4.25mmoj2) Z — Lys (Boc) — N — (3-pyridylmethyl) amidine is dissolved in 20m5TFA 'solution at 0 ° C and stirred for 20 minutes. The excess TFA is concentrated and oily residue. This paper is in accordance with Chinese national standards ( CNS) A4 specification (2I0X297 mm) -44-A 7 ——— _; _ · __ 5. · Description of the invention (42) The product is treated with 10mj? DMF. Then 4 · 6mj? (42. 5mm.) i2) N-methylmorpholine, 1.15 g (4.25 mm)? 4-[[4-aminoiminomethyl) phenyl] amino] -4-oxybutyrate , 2.35g (5.3m m $) Bop and 20m $ DMF. The mixture was stirred at room temperature for 24 hours. The DMF was concentrated, the residue was extracted twice with 4 OmJ? Water, filtered with suction and dried. The crude product was purified by chromatography on silica gel using an eluent 89b (70 mole% HCC 芡 3 per liter of NH4OH 25%, 40 mole %% 6011, 10 mole% CH3C00_Na +). Yield: 34 〇mg (14% of the theoretical value). Examples 36 to 55 were obtained in the same manner as in Example 35. (Please read the precautions on the back and fill in the "quotation page"), τ.¾ Central Standard of the Ministry of Economic Affairs Printed by Zhang Shi M S C / (\ j quasi-standard one country, one country, one country, one use, one centimeter 7 9 2 A7 B7 V. Description of the invention (43) Table 4: Melting points of the compounds according to Examples 3 5 to 5 5 Example mp [° C] Example mp [° C] Example mp [° C] 35 190-198 42 190 49 189 36 218-220 43 198 50 197 37 209 44 213 51 38 195 45 52 39 189-191 46 175 53 40 215-220 47 196 54 194 41 183 48 217 55 ------------- install -------- order --------- line (please read first Note on the back side 'Please fill in this page again) This paper size applies to Chinese national standard (CNS> A4 size (210 x 297 mm) -46-A7 B7

Read the notes on the back of it before f

Printed by A7 _ _B7___ of the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (45) 1. Use the carboxylic acid or chloroformate for the tritiation according to the schematic diagrams 4 and 5: H — Ly s (Bo c) — NH2 It is reacted with a carboxylic acid in an aprotic solvent (DMF, DMSO) in the presence of a base (DI PE A, NMM) and a coupling reagent (DCC, DI C, EDCI) at room temperature to produce the formed amidine. After removing the solvent, the residue was treated with water, and the less insoluble crude product was filtered by suction. The product can be purified by crystallization from alcohol (MeOH, EtOH < 2-PrOH) or ester (MEK, EA). "H-Lys (Boc)-NH2 and carbonyl chloride in alkaline aqueous solution (Schotten-Baumann conditions) The reaction yielded the desired derivative in a yield of 90-95%. Isolate the crude product by suction filtration and purify β by recrystallization from alcohol (Me OH / E t 0H / isopropanol) or ethyl acetate or methyl ethyl ketone. 2. Remove Boc protecting group using TFA. Removal of the Boc protection group in a mixture of dichloromethane and trifluoroacetic acid (2: 1) at room temperature is achieved after about 30 minutes. Please read the precautions on the back and then% [本 ' page

Order I quantitative. The residue Ri_L y, which is usually oily after isolation, reacts quickly without further purification steps. N Η 3. Tritiated, in which R4 = 4- ((4-aminoiminomethyl) phenyl) amino) -4 monooxybutyrate: Reaction with other carboxylic acids (R4) At room temperature, in the presence of alkali (this paper size applies Chinese National Standard (CNS) A4 specifications (210X297 mm) 48 ~ A7 ______B7 _ V. Description of the invention (46) nMm, DIPEA), using such as EDCI, B OP or PyBop was performed in an aprotic solvent (DMF'DMSO). After the solvent was removed, the product was precipitated when water was added. Purification was performed on a R P 18_ column using a mixture of eluent of water, acetonitrile, and trifluoroacetic acid 'by HP HP. The product was obtained in the form of a TFA salt. According to this general method (the basis of the synthetic schemes 4 and 5), compounds were synthesized as described in the following Examples 5 6 and T Table 5: At room temperature, 3 2 mm 〇j? Z-lysamine amine hydrochloride And .2 2 mm 〇 ^ 4- ((4- (aminoiminomethyl) phenyl) amino) -4-oxybutyrate is added to 1 2 Omj? Anhydrous and degassed N, N-dimethylformamide (DMF). The starting material was quickly dissolved under stirring; after adding 104 mm of diisopropylethylamine and 40 mm of Bop, the mixture was stirred at rt for 16 hours. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) I "! ^ On a rotary evaporator, at a bath temperature of 5 0 _ 5 5 ° C and about 1 0 Under mb ar, remove solvent and excess DI PEA. The oily residue was treated with 2 5 Omj? Water, homogenized in an ultrasonic bath, and cooled. The precipitated crude product was filtered off by suction and washed with water on a suction filter. After drying on calcium chloride in vacuum, about 16 g of HCJ2 salt form of a beige powder were obtained with a purity of about 90% (HPLC). When preparing the corresponding trifluoroacetate, the product is suspended in 100MJ2 water, and 32mmoj? (2.45m meaning) trifluoroacetic acid (99 paper size applies to Chinese National Standard (CNS) A4 specifications (210 '/ 297) Printed for implementation by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs! R1-CO r2 / r3 R4 56 ^ '0 H / H 57 〇58 • 59 Spring paper size is applicable to the Chinese National Standard (CMS) A4 specification (210X297 mm) ) A7 B7 V. Description of invention (47)%). In order to remove the excess acid 'mixture again on a rotary evaporator, a short vacuum was applied, and then the aqueous suspension was freeze-dried. After recrystallization from alcohol (EtOH / MeOH), the resulting product was freeze-dried again to obtain better solubility. Yield: 5. 2 6 g

M.p .: 210-213 ° C R2

R1-CO-NH-CH-CO-N I ''

I

NH Γ CO-R4 (Formula I) Table 5: a, e—N—Substituted L-lysamidamine derivatives according to the schematic diagrams 4 and 5 (where all examples are equal to 4)

I -50-Example 1 printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs! Γ + R1-CO r2 / r3 R4 79 H / H ^ -CH: 80 Clv ^ 〇 ^ 〇81 82 A7 B7 V. Description of the invention ( 48) Table _ (continued) This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling in this page)

A7 B7

V. Description of the invention (Form 49 __ (Continued) Real._J

R-CO R2 / R3 60

H / H

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 61 、 0〆 _! 62 Γ ^ ί 0 63 〇 64 65 (\ Cr. 0 66 c-XJ 67 Op 68

This paper size applies to China National Standard (CNS) A4 (210X 297 mm) (Please read the precautions on the back before filling this page)

7 7 A B V. Description of the invention (50) Form __5_ (continued) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

(Please read the precautions on the back before filling in this page) # 纸 尽 A \ J / s N c ί Biaozuo Guoguo Tongtong X ο Centimeter Attachment c: No. 85114601 Patent Application Chinese Specification Revision Page Republic of China 88 Presented in September 2015. Description of the invention (51) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs, Table 6: Melting point of the compound according to Example 5 6 to 8 Example m. P. [° C] Example m, p. [° C] Examples m. p, [° C] 56 210-213 65 218-222 74 191-193 57 220-223 66 216-219 75 186-188 58 213-215 67 235-238 76 220-222 59 223-226 68 Gundam 218 77 210-215 60 Gundam 233 69 205-208 78 Gundam 223 6 1 Gundam 237 70 168-170 79 Gundam 226 62 Gundam 221 71 197-202 80 194-197 63 Gundam 220 72 22 Bu 226 81 215-222 64 230-236 73 225-228 82 219-222 (Please read the precautions on the back before filling out this page]-丨 Installing _________ 泉 · 丨 This paper size applies Chinese national standards ( CNS) A4 specification (210 X 297 mm) -54-.A7. _B7 _ ^ ___ V. Description of the invention (52) Note: ^ up to ... means that the substance is frozen Dry (please read the notes on the back before filling in this education) After drying, it will form an amorphous foam with corresponding physical properties. The narrow melting point does not exist, but it is slowly sintered to liquefaction. The compounds of the invention may also be in the form of acid addition salts, such as inorganic acid salt forms, such as, for example, hydrochloric acid, sulfuric acid, phosphoric acid, organic acid salt forms * such as, for example, acetic acid, trifluoroacetic acid, lactic acid, malonic acid, maleic acid, Fumaric acid, gluconic acid, glucuronic acid 'citric acid' Inbernic acid, methanesulfonic acid, isethionic acid, pyruvate and succinic acid. Both compounds of the general formula I and their salts All are biologically active. The compounds of the general formula I can be administered in free form or in the form of a salt formed by a physiologically tolerable acid. The medication can be administered orally, parenterally, intravenously, transdermally or by inhalation Business.-

Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. The present invention also relates to a pharmaceutical formulation containing at least one compound of formula 1 or a salt formed with a physiologically tolerable inorganic or organic acid. Excipients and / or diluents or adjuvants can be used. Example 8 3 Cetrorelix, Example 1, Example 2 and Example 5 6 Bonding affinity for human LH-RH receptor. (West Torres: Ac — D — Na 1 (2) — D — ρ — Cl — Phe-D-Pa 1 (3)-Se r-Tyr-D The paper dimensions apply to the Chinese National Standard (CNS) A4 Specifications (21 OX29 ^ mm) A7 printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs —— ^ _; _- V. Description of the invention (53) C it— Leu — Arg — Pro— D — A1 a — NH 2 determination Method of Bonding Affinity (Dissociation Constant Kd): Determination of Bonding Affinity by Competitive Bonding Test (A substitution bonding test; Beckers et al., Eur. J. · Biochem. 2.3 1 .., 5 3 5 1 543, 1995). The radiolabeled ligand used was [12SI] Citorex (specific activity 5-1 0 X 1 05d pm / 1; dissolved in 20% v: v acetonitrile, 0 ... 2 % w :. v protein '0.1% w: v TFA, ~ 80% v: v aqueous solution). The bonding capacity of the iodinated peptide is between 60% and 85%. It is not used. The test compounds are Citorex, solution 1, Example 2 and Example 5. The use concentration of the substance is 〇1 〇1 nM -1 OOOnM (Citorex, Example 1, Example 1). Example 2) or 0.011 to 1 ^ -1 to 10) Example 5 6). 'Individual cells fine L 3. 5/7 8 used on the surface of human LH- RH receptor overexposure in the bond test are PBS / EDTA (Ca2 + + / Mg2 + / lmM EDTA free PBS) was removed from the culture dish cultured under non-fusion conditions, and the cell count was measured. The cells were resuspended in the incubation medium at the corresponding cell density (Dulbecco's modified Eagle medium contains 4.5 g / ^ glucose, 10mM Hepes _ρΗ7.5 '0.5% w ·· v BAS., lg / j? bacitracin, 0.1 g / 芡 SBTI, 0.1% w: v NaN3). Firstly 200 00 $ Silicone / paraffinic oil mixture (84/1 6 vol%) The paper size of this paper applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) (Please read the precautions on the back first, fill in this, buy this) 、 1T ------ line # · Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 ____B7__ V. Description of the invention (54) Into a specific 400 Renren reactor (1? 611116): 36 111311), and aspirated cell suspensions (2.5 x 105 cells) were placed on top of each other. To the cell suspension above the silicone / paraffinic oil layer was added a 50 $ bond medium containing the appropriate concentration of [125 I] Citorex and the compound to be tested. The mixture was spun in a greenhouse at 37 ° C for 60 minutes. After this step, the cells were pelleted in a silicone / paraffinic oil layer during centrifugation in ΗTA13.8 rotor in Heraeus Biofugel 5 for 2 minutes at 9000 r pm (at room temperature), and Separate from bonding medium. After centrifugation, the reactor was quick-frozen in liquid N2. Using a pair of forceps, the reactor tip (cell pellets) was cut off. The tip of the liquid (unbound free ligand [1251] Citorex) was moved to the counting tube. When the maximum bond value (B 0) was measured, no competitor was added. When measuring non-specific binding, 1 β M unlabeled citriurex was added to facilitate competition. When the total S bond is 10%, the non-specific bond is low. Quantification was performed in a 7-counter; analysis was performed using the EBDA / ligand V3.0 program (Mcpherson, J. Pharma c'ol. Methods 14, 213-228, 1985). Plotting in a dose-response plot estimates I C 5. (Concentration that causes 50% inhibition of the receptor response), and the EBDA / ligand program calculates the dissociation constant Kd (nM) °. Results: According to the competition curve (see Figure 1), all test compounds are shown to bond with The radiolabeled ligand [1 2 5 I] that competes with the human LH-RH receptor competes for Citoralis. In each case, the Chinese National Standard (CNS) A4 specification (210 × 297 mm) applies to the paper size of the competition: -57----: ------, I ', > ------ -Order ------ line ^ 1] --- ΊΊ. „. '., _ + (Please read the notes on the back before filling in this page) A7 ___.__ B7 _________ V. Description of the invention (55) The concentration of the agent plots the bondability (expressed as% of total bondability Bo). For the compound shown in 1, the following bond affinity can be calculated in terms of the dissociation constant K d [nM]: Citorex ( SB-75)-214 nM, Example 1-0. 305 nM 'Example 2-0. 10 4 nM, and Example 56-98 6 nM. The bond affinity of the average value of each measured value can be obtained from Table .7 Example 8 4 Example 2 and Example 5 6 Antagonism of human ^ L H_R Η receptor in functional group assay Ip 3 (D-my ο-1,3,5-triphosphate Ester method: Subculture of human LH-RH receptor overexpressing cell line (L3. 5/78 printed by the Employees 'Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs), washing the culture with PBS, and removing the cells with PBS' / EDTA To suspend cells "Pellets formation" cells were resuspended in incubation medium (Dulbecco's modified Iger medium, containing 4.5g / j? Glucose, 10nM Hepes pH 7.5, 0.5% w: v. BSA, 5mM LiCj ?, lg / Coclopeptide, 0.1g / 5 SBTI), aliquoted in a 1.5mS reactor, and pre-incubated for 30 minutes at 37 ° C. Each measurement point needs 4X1 〇β cells in a 500 # volume After the pre-incubation step, a final concentration of 10 nM LH-RH (a stock solution at 10 mM Tris pH 7.5, lmM dithiothreitol, 0.1% w: v BSA / Bachem Art # 4 0 was added to the cell suspension. 0 5 in 0.5 mM). By the same paper size as the Chinese National Standard (CNS) A4 specification (21 OX297 mm) -58-Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (56) Add the corresponding concentration (such as 0.0316, 0.1, 0.316, etc., examples up to 100nm) to test the effect of the antagonist. The negative control group was cultured without LH-RH. The cells were incubated at 37 ° C. After 15 minutes, trichloroacetic acid (TCA) was used to extract the IP 3 formed from the cells isolated. Finally, in the cell suspension 5 Ο β 5 was added to the ice-cold 15% (w: v) TCA solution to form a precipitate. The material was formed by centrifugation in a Heraeus Biofage 1 5 R centrifuge at 4 '° C for 15 minutes at 2000 xg for 15 minutes. Pills. 950 # 芡 Supernatant was extracted in a 15mj? Container (placed on ice) three times with a volume of 3 x 10 times saturated with water-saturated cold diethyl ether. After the final extraction step, the solution was adjusted with 0.5M NaHC03 solution to pH7.5. The concentration of I P 3 in the cell extract was determined by using the I 3 -binding protein, labeled [3H] -I P3, and unlabeled I P3 by a competitive competitive bonding test. Finally, use Amersham's test kit (TRK 1 00 0); perform the measurement as described in the test method. After various steps were performed, 2 m J of a scintillation counter (Rotiszint Ecoplus) for a water-containing sample was finally added, and the pellets containing bonded [3 Η] -IP 3 were carefully re-suspended after being 'resuspended', And in a scintillation counter, the cell IP 3 content was calculated using a standard curve, and a dose-response curve was set. I C50 is estimated from the inflection point of the curve. Results: Figure 1 shows a dose-response curve suitable for peptide antagonist example 2 (Figure 2), and peptidomimetic example 56 (Figure 3). Stimulation was performed with Γ0 η M LH-RH, and the inhibitory ability to form I P3 was measured as a function of the concentration of the substance. As far as Example 2. and Example 5 and 6 are concerned, it is impossible to determine the size of this paper. Applicable to China National Standard (CNS) A4 specification (210X297 mm) (57) Any antagonistic activity, that is, the substance itself cannot cause any stimulation to IP 3 synthesis. The IP 3 concentration that was still measurable at the highest concentration corresponded to unstimulated cells. In Examples 2 and 56, we dealt with LH-RH functional antagonists. However, the effectiveness of this substance is different. Under the selected experimental conditions, I C5 of Example 2. It is about 0.4 nM, and IC 5 of treasure example 5 6. Approximately 4 μM. These activities are closely related to the in vitro bonding affinity, which was determined in a competitive bonding test using [1 2 5 I] -citorex, Kd = 0.109 nM in Example 2, Example 56 Kd = 1.0 8 / ζM »Example 8 5 Example 1, Example 2 and Example 5 6 Hormone Inhibition in Healthy Male Rats When the inhibition of fluorenone is measured in the blood of healthy male rats' The substance was injected into the right lumbar fossa of the animal. The dosages of Examples 1 and 2 were 1.5 mg / kg, while that of Example 56 was 1 Qmg / kg. When the fluorenone value was measured, about 300 m of animal blood was taken from the sublingual vein at 0.2, 4.8 (example 56 only) '24, 48, 72, and 96 hours', and then taken every three days until the suppression was stopped. The drug of Example 1 was used to inhibit 1'g / ιηβ fluorenone for '2.64 hours in one animal, 3 3 6 hours in two animals' and 3 8 4 hours in one animal (Figure 4) ). After taking the drug of Example 2, the concentration of fluorenone in one animal was suppressed for 408 hours and 648 hours in four animals (Figure 5). Example 56 (10mg / kg M-scale scale is applicable to Chinese National Standard (CNS) A4 specification (2 丨 OX 297mm) (Please read the precautions on the back before filling in this page). One, -ιτ --- -Line i · — ^ — • im Hal nn nn -60-Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 __B7_ V. Description of the invention (58) s.c.) Even after 2 hours, all 5 The concentration of fluorenone in the animals was maintained for 8 hours. At subsequent measurement points (24 hours). The fluorenone value increased again (Figure 6). Table 7: Bonding affinity of biological data to human L Η-RH receptors (expressed as dissociation constant Kd [ηM]; evaluated using EBDA / ligand analysis program. Mean values from various experiments are shown, and the experiments are in parentheses Times) and acetone inhibition in vivo, histamine release in vivo, and water solubility compared with S Β-7 5: This paper size applies Chinese National Standard (CNS) A4 specification (210 × 297 mm) (Please read the precautions on the back before (Fill in this page)

-61-A7 B7 V. Explanation of the Invention (Others) Printed Substance Affinity Human LH-RH Receptor [nmo1 / L] (1.5mg / kg, single dose) by the Central Bureau of Standards, Ministry of Economic Affairs, Inhibition in Rats [ h] (IC 5 0) Histamine release [β g / ml] h2〇 Solubility [mg / m1] Citorex s Β-7 5 0.202 (10) 144 9.7 9 Example 1 0.306 (2) 336 31. 9 27 Example 2 0. 109 (2) 648 17. 1 23 Example 3 0.170 (2) 864 nd * nd Example 4 0.206 (2) 696 η. Dnd Example 5 6 1082 (2)-- -$) Unable to measure due to poor solubility. (Please read the notes on the back before filling in the 'write this page') The size of the paper used in this edition applies to the Chinese National Standard (CNS) Α4 specification (210X297 mm). _ Printed and repaired on the A 7 Leap Year Month Day B7-3-1 5 ————————- V. Description of the Invention (60—) Passage Jane · Explanation Figure 1 shows Cetrorelix, Example 1 Competitive curves for the binding of human, L, and RH receptors by compounds of 2, 2, and 56. Figure 2 shows the antagonistic effect of the compound of Example 2, which is a cell line that is overexpressed by the human RH and RH receptors (L3. 5/78) The IP 3 concentration in the extract was measured. Figure 3 shows the antagonism of the compound of Example 56 as measured by the concentration of IP3 in the extract of a cell line (L3.5 / 78) overexpressing the human LH-RH receptor. Figure 4 shows the inhibition of fluorenone in the blood of rats after injection of the compound of Example 1. Fig. 5 shows the inhibition of fluorenone in the blood of rats after injection of the compound of Example 2. Figure 6 shows the inhibition of fluorenone in blood after rats were injected with the compound of Example 56. The paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page) Ding ______: Yuan loses ________ -'aL2 ^ • 63-

Claims (1)

Announcement A8 B8 C8 D8 58 · 9, ί 3 See {f · year month 丨 less one '. One. Supplementary Patent Application Scope Annex AI No. 8 5 1 1 4 6 0 1 Chinese Patent Application Scope Amendment Amended in September 1988 1. A compound of general formula (I), R1-C〇-NH-CH C0- (f2) 4 NH CO-R4 -N, R (I) (Please read the note on the back first Please fill in this page for further information.) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, where R1 is Ci-C6 焼 '' C i-C alkoxy, which is optionally substituted with halogen, I Ce-C i 2 aryl, which Arbitrarily substituted with halogen, quinomethoxy or phenylferroyl, Ce—C12aryl—Cl—Ce alkyl, which is optionally substituted with halogen, Ce Ceoxy or phenyl * Ce—C12aryl — (^ ace Alkoxy, which is optionally substituted by nitro and R 2 and R 3 are each a hydrogen atom, C 1 ~~ C β alkyl, phenyl-Cl- ce alkyl, which is optionally amine, halogen or (^- c β alkoxy substituted, heteroaryl-Ci-ce alkyl, wherein the heteroaryl is selected from pyridyl, hexahydropyridyl, furanyl, imidazoben & Zhang scale applicable to China Standard (milk milk) point 4 specifications (210 fathers, 297 males) _1_ " Announcement A8 B8 C8 D8 58 · 9, ί 3 See {f. 8 5 1 1 4 6 0 Chinese Patent Application No. 1 Amendments to the Republic of China 8 September 1998 Amendment 1. A compound of general formula (I), R1-C〇-NH-CH C0- (f2) 4 NH CO—R4 -N, R (I) (Please read the notes on the back before filling this page) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs where R1 is Ci-C6 焼 '' C i — C le alkoxy Group, which is optionally substituted with halogen, I Ce—C i 2 aryl group, which is optionally substituted with halogen, quinomethoxy or phenylferroyl group, Ce—C12 aromatic-Cl-Ce alkyl group, which is optionally substituted with halogen, Ce 焼 oxy or phenyl substituted * Ce- C12 aryl — (^-ce alkoxy, which is optionally substituted by nitro and R 2 and R 3 are each a hydrogen atom, C 1 ~~ C β radical, phenyl A Cl-ce alkyl group, which is optionally substituted with an amine group, a halogen or (^ -c β alkoxy group, a heteroaryl-Ci-ce alkyl group, wherein the heteroaryl group is selected from pyridyl, hexahydropyridine , Furanyl, Imidazoben & Zhang scales are applicable to Chinese National Standard (标准 奶) spot 4 specifications (210 fathers 297 males) _1_ " VI. Application for patent scope base, evil well base, benzimidazolyl and pyrrolidine oxidation And this heteroaryl group is optionally substituted by Cl-ce alkyl; or -NR 2R 3 is an amino acid group; R 4 is a group of the general formula (II)-(CH2) p-CO-NR5R6 (id where P mil is an integer of 1 to 4, R-5 is hydrogen, and R6 is C. "Aryl" This aryl group is optionally methyl, cyano, halogen, ca-Ce alkyl, Cl-fluorenyloxy, benzyloxy, trifluoromethyl or (: 1-(: 4 alkoxycarbonyl) Ethyl is substituted or is a heteroaryl group selected from pyridyl, pyridyl, and pyrimidinyl 'wherein the pyridyl is optionally substituted with halogen, amine, or Ci-alkoxycarbonyl; or R 4 is Ring of general formula (III) N) = 〇N (III). Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page). The palm carbon atoms can have the R- or s-configuration 'and It is a salt with a pharmaceutically acceptable acid. 2. The compound of item 1 in the scope of the patent application, which is selected from α-N — Z — [ε — — 4 — (4-Phenyl) amino-1 , 4-dioxybutyl] lysamide and its salts with pharmaceutically acceptable acids. .0X1. 3. If the compound of the scope of patent application No. 1 is selected from the paper standard applicable to the Chinese National Standard (CNS) A4 specification (210x297 thin) Printed by A8 B8 C8 D8 of the Consumer Cooperatives of the Central Bureau of Standards • Scope of patent application: N—Z— [ε—N—4— (4-Fatphenyl) amino-1,5-dioxypentyl] lysamide and its pharmaceutically acceptable acid Into the salt. 4. The compound according to item 1 of the scope of patent application, which is selected from the group consisting of α-N-Z- [e-N <-( imidazolidine-2-ketone-4yl) -methylamidinyl] lysamide and Salts with pharmaceutically acceptable acids. 5. The compound according to any one of claims 1 to 4, wherein the salt is erabonate »6. A compound of general formula (V), Ac-D-Na 1 (2) 1— D (pCl) Phe2- D — P a 1 (3) 3 — S er 4 — T y r5— D — Xxx6 — Le eu 7 — A rg 8 — P ro 9 — D — A 1 a 10 — NH 2 (Please read the precautions on the back before filling in this page) Equipment, 1T V where D_Xxx is the amino acid group of general formula (VI) • NH—CH—C0- CH 2, NH CO——R (VI) wherein R 4 is a group of the formula (II)-(CH2) p-CO-NR5R6 (II) wherein p is an integer from 1 to 4, R 5 is hydrogen, and rule 6 is 06-: 1. Aryl group, this aryl group is arbitrarily methyl, cyano, halo ^ 'C! — C 6 alkyl, this paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) 6. Apply for patent n______________ Cl — ce fluorenyl, benzyloxy, trifluoromethyl, or Cl — c4 alkoxycarbonylethynyl, or a heterocyclyl selected from pyridyl, pyridyl, and pyrimidinyl, where the pyridyl Optionally substituted with halogen, amine or Ci-c4 alkoxycarbonyl; or R 4 is ring X of the general formula (III) = 0 N (III) and a salt thereof with a pharmaceutically acceptable acid. 7. The compound according to item 6 of the scope of patent application, wherein Xxx is, [ε-N-4-(4- Fatty Phenyl) amino-1,4-dioxybutyl] pyrimidine. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) 8_ If the compound in the scope of patent application No. 6 is applied, where χχ is [ε-N-(imidazolidine-2-one -4-yl) methylfluorenyl] lysaminomethyl. 9. The compound according to any one of claims 6 to 8, wherein the salt is an embonate. I. A pharmaceutical composition for use as an LH-RH (luteinizing hormone-releasing hormone) antagonist, including a compound according to any one of claims 1 to 9 of the patent application scope. II. The size of the paper used to prepare the compound in item 6 of the scope of the patent application is subject to the Chinese National Standard (CNS) A4 specification (210X297 mm). 6. The scope of patent application method, which includes the following steps: (a) in D-Lai Provide appropriate protective groups on the α-amine and carboxylic acid groups of the amino acid or D-ornithine, (b) make the D-lysine or D-ornithine with the protective group and the carboxylic acid of the general formula (VII) Acid reaction R 4-C Ο Ο Η (VI I) where R 4 is as defined in the first patent application scope, (c) removing the protecting group on the α-carboxylic acid group of the compound obtained in step (b), and In step (h), position 6 is incorporated, (d) the D-alanine having a protective group on the amine group is coupled to the solid support of the resin type, and (e) the protection on the amine group of alanine is removed (F) reacting alanine bonded to a solid support with proline having a protective group on the nitrogen atom, (g) removing the protective group located on the nitrogen atom of proline, Ministry of Economic Affairs Printed by Lingyang Standard Bureau staff consumer cooperatives (please read the precautions on the back before filling this page) (h) Use steps c) The modified D-lysine or D-ornithine described in position 6 for amino acid 1 to 8 of the general formula (V) is repeated according to the order from 8 to 1 And g), (i) remove the compound obtained in step (h) from the monomer and purify it if necessary, especially using HPLC, (j) if necessary, react with a pharmaceutically acceptable acid to cause Bernic acid is preferred. 1 2. A method for preparing a compound as described in item 6 of the scope of patent application, which includes the following steps: This paper size applies the Chinese national standard (CNS > A4 specification (210X297 mm)) 6. The scope of patent application (a) D-alanine with a protective group on the amine group is coupled to a support suitable for solid-phase synthesis, (b) removing the protective group on the amine group of alanine, (c) bonding to the resin Alanine reacts with proline having a protecting group on the nitrogen atom, (d) removing the protecting group on the nitrogen atom of the proline, (e) using amino acids 1 to 8 of the general formula (V), Repeat steps c) and d) in the order of 8 to 1. (f) Remove the compound obtained in step (e) from the support. (G) react with the carboxylic acid of formula (VII). R 4-C 〇 〇 Η (VI I) where R 4 is as defined in item 1 of the scope of patent application, and (h) if necessary, it will react with a pharmaceutically acceptable acid, preferably Inboic acid. 1 3. — A method for preparing a compound such as the scope of patent application No. 6, including the following steps: Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) (a) D-alanine having a protective group on the amino acid is coupled to a support suitable for solid-phase synthesis, (b) removing the protective group on the amino group of alanine, and (c) bonding to the resin The alanine above reacts with proline having a protecting group on the nitrogen atom, (d) removes the protecting group located on the nitrogen atom of the proline, (e) uses the amino acid of formula (V) 6 to 8 Repeat steps c) and d) according to the order from 8 to 6. This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 cm). 6. The scope of patent application (f) is from D-lysine or D_Ornithine removes the e-amino protecting group at position 6 and reacts with the carboxylic acid of formula (VII), R4-COOH (VII) where R 4 is as defined in the first item of the patent application scope, ( g) removing the protective group on the α-amino group of D-lysine or D-ornithine, (h) using the amino group of general formula (ν) 1 to 5, in the order of 5 to 1, repeat steps c) and d) e (i) remove the compound obtained in step (h) from the resin and purify it, especially using ΗPLC, (j) if necessary , It reacts with a pharmaceutically acceptable acid, preferably inbornyl acid. 14. The method 1 according to any one of claims 11 to 13 in the scope of patent application, wherein N— (4-fatphenyl) amino-4—oxybutyric acid is used as the formula (VII) carboxylic acid. 15. The method according to any one of claims 11 to 13 in the scope of patent application, wherein imidazolidine 2-keto-4 monocarboxylic acid is used as the general formula (printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs VII) Carboxylic acid · 16. The method according to any one of claims 11 to 13, in which inbornyl acid is used as a pharmaceutically acceptable acid. 17. The compound according to any one of claims 1, 2, 3, 4 or 6 *, which is used for the manufacture of hormone-related tumors, especially prostate or breast cancer, and the supply and demand inhibition of LH-RH Hormonal therapy for non-malignant tumor indication drugs * This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling this page) ----