CN1017436B - Peptide containing immunoexcitant - Google Patents

Abstract

The present invention relates to peptide components of formula 1, pharmaceutically acceptable base salts thereof, pharmaceutical compositions and their use as antiinfective agents where R1 is alkyl, cycloalkyl or cycloalkylmethyl; R2 is hydrogen or alkyl and R3 is hydroxy or an amino acid residu of the formula where X is hydrogen, alkyl or hydroxymethyl and n is an integer of 0 to 4 and R4 and R5 are alkyl, hydrogen, benzyl or cyclohexylmethyl.

Description

Peptide containing immunoexcitant

The present invention relates to as the novel acyl glutamic acid that contains peptide class immunostimulant and anti-infection agent; Their formula of medicine and their purposes aspect the treatment infection.

Immunopharmacology than frontier, especially adjust this minute field handling immunity, continue advancing with fast paces.People once studied various natural compounds, and comprising tetrapeptide Ta Fuxin (tetrapeptide tuftsin), known its chemical formula is N ²-(1-N ²-L-threonyl-L-lysyl)-the L-prolyl)-the L-arginine.Many people notice the synthesize polypeptide sugar derivatives; Especially two peptide classes of those known muramyls.Adjust agent, especially a large amount of compounds of studying as immunostimulant in order to sum up as immunity, people can be with reference to following works: Duker etc., Annu, Rep.Med.Chem., 14,146～167(1979), Lederer, J.Med.Chem., 23,819～825(1980) and J.Kralovec, Drugs of the Future, 8,615～638(1983).

The excited peptide class of immunity was once described in some patent specifications:

The two peptide classes of L-alanyl-α-glutamic acid N-acyl group are stated from the Deutsches Reichs-Patent of announcing on January 15th, 1,981 3,024,355;

The tetrapeptide class and the pentapeptide class that contain D-alanyl-L-glutamyl part or L-alanyl-D-glutamyl part are stated from English Patent of announcing on February 4th, 1,981 2,053,231 and the Deutsches Reichs-Patent of announcing on January 8th, 1,981 3,024,281 respectively; And N-acyl group-alanyl-γ-D-glutamyl tripeptide derivative; wherein the C-end amino acid is Methionin or diaminopimelic acid; be stated from the Deutsches Reichs-Patent of announcing on January 15th, 1,981 3; 024; 369, and the lactoyl tetrapeptide class that disclosed European patent 11283 was made up of N-lactoyl alanyl, glutamyl, diamino pimeloyl and carboxymethylamino component on May 23rd, 1980.

Other also has, and has the excited polypeptide class and the derivative thereof of immunity of following formula (A), is stated from United States Patent (USP) 4,311,640 and 4,322,341; Europe number of patent application 25,482; 50,856; 51,812; 53,388; 55,846; With 57,419.

R in the formula (A) ¹Be hydrogen or acyl group; R ²Hydrogen particularly, low alkyl group, methylol, benzyl; R ³And R ⁴The hydrogen of respectively doing for oneself, carboxyl-CONR ⁷R ⁸, R wherein ⁷Be hydrogen, can be by the low alkyl group of hydroxyl replacement; R ⁸Be mono carboxylic low alkyl group, dicarboxyl low alkyl group; R ⁵For hydrogen or for carboxyl, its condition be to work as R ⁴And R ⁵The two one of be hydrogen, another be carboxyl or-CONR ⁷R ⁸; R ⁶Be hydrogen; M is 1 to 3, and n is 0 to 2, with and derivative, wherein carboxyl and amino are protected.

Disclosed aforementioned polypeptides class in the prior art is not also at the variable R of following formula ⁴Contain the heterocyclic radical part on the shared position, the polypeptide class of in the Application No. 595,169 of the Application No. 662,668 of on October 19th, 1984 application and application on March 30th, 1984, describing by people such as Ives only, wherein variable R ⁴It is a kind of alkali type amino acid part.

Kitaura etc., at J.Med.Chem., reported the minimal structure N of the biological respinse characteristic that can cause formula (A) compound on 25 335～337(1982) ²-(γ-D-glutamyl)-meso-2(L), 2(D)-diaminopimelic acid; N is 1 in the formula (A); R ¹Be CH ₃CH(OH)-CO-; R ²Be CH ₃; R ³And R ⁵Respectively do for oneself-COOH; R ⁴For-CONHCH ₂COOH; R ⁶Be H.Said above-mentioned formula (A) compound is called as FK-156.

Novel immunostimulant of the present invention is to have the compound of following formula 1 and can be used for pharmaceutically subsalt,

Wherein, R ₁It is the alkyl that contains the cycloalkyl of 4 to 7 carbon atoms or contain 2 to 10 carbon atoms; R ₂For hydrogen or contain the alkyl of 1 to 3 carbon atom; R ₃For hydroxyl or have the amino-acid residue of following formula

Wherein, X is a hydrogen, contains the alkyl or the methylol of 1 to 2 carbon atom, and n is 0 to 4 integer; R ₄And R ₅The hydrogen of respectively doing for oneself contains the alkyl of 1 to 6 carbon atom, contains the cycloalkanes methyl or the benzyl of 6 to 8 carbon atoms.

One group is those compounds through preferred compound, wherein, and R ₁For containing the alkyl of 5 to 8 carbon atoms, R ₂Be hydrogen, R ₃Be above-mentioned amino-acid residue, wherein, X, n and R ₅As defined above, R ₄Be hydrogen.Particularly preferably be those compounds in this group, their n is 0, R ₅Be hydrogen, said alkyl or cyclohexyl methyl.Particularly preferred compound is those compounds, wherein, and R ₁Be (R, S) 2-ethyl-1-butyl, R ₅For hydrogen, X are methyl; R ₁Be (R, S) 3-heptyl, R ₅For hydrogen and X are methyl; R ₁Be (R, S) 2-methyl-1-pentene base, R ₅For hydrogen and X are methyl; R ₁Be (R, S) 2-heptyl, R ₅For hydrogen and X are methyl; R ₁Be (R, S) 2-ethyl-1-amyl group, R ₅For hydrogen and X are methyl; R ₁Be (R, S) 1-hexyl, R ₅For hydrogen and X are methyl; R ₁Be (R, S) 2-ethyl-1-hexyl, R ₅For hydrogen and X are methyl; R ₁Be (S) or (R, S) 2-methyl isophthalic acid-hexyl, R ₅For hydrogen and X are methyl; R ₁For (S) or (R, S) 2-ethyl-1-hexyl, X are methyl and R ₅Be hydrogen and R ₁Be the 1-hexyl, X is methyl and R ₅Be hydrogen.R wherein ₁Being the 1-hexyl, when X is hydrogen and n=3, also is particularly preferred compound.Especially preferred ester is those esters, wherein R ₁For (R, S) 2-ethyl-1-amyl group, X are methyl and R ₅Be normal-butyl, isobutyl-or cyclohexyl methyl; R ₁Be (S) or (R, S) 2-methyl isophthalic acid-hexyl, X are methyl and R ₅Be normal-butyl, isobutyl-or cyclohexyl methyl, and R ₁For (S) or (R, S) 2-ethyl-1-hexyl, X are methyl and R ₅Be normal-butyl, isobutyl-or cyclohexyl methyl.

Second group of preferred compound is those compounds, wherein, and R ₁Be the cycloalkyl that contains 4 to 7 carbon atoms, R ₂Be hydrogen, R ₃Be above-mentioned amino-acid residue, wherein n is 0, and X is the alkyl that contains 1 to 2 carbon atom, R ₄And R ₅The hydrogen of respectively doing for oneself.In this group, particularly preferably be those wherein R ₁For cyclohexyl and X are the compound of methyl.

The 3rd group of preferred compound is those compounds, wherein R ₁The alkyl that contains 5 to 8 carbon atoms, R ₂Be hydrogen, R ₃Be above-mentioned amino-acid residue wherein X be hydrogen or the alkyl that contains 1 to 2 carbon atom, n is 0 to 4 integer, R ₅Be hydrogen, R ₄For containing the alkyl of 1 to 6 carbon atom, contain the methyl cycloalkyl or the benzyl of 6 to 8 carbon atoms.

The present invention is also at the formula of medicine of unit dosage, and this prescription comprises the carrier that is suitable for doing medicine and as formula 1 compound of anti-infection agent or immunostimulant significant quantity; And the method for the treatment of infected patient, this method comprises that the people who is infected takes formula 1 compound of anti-infective dosage.

The subsalt that can be used for medicine of said formula 1 compound is meant the salt of those mineral alkalis or organic bases, as alkali metal hydroxide and alkaline earth metal hydroxides, and the salt of ammonia, triethylamine, thanomin and dicyclohexyl amine.

The configuration of the amino acid moiety of composition formula 1 compound is important with regard to the pharmacologically active of said compound.In having specified stereochemical those compounds of formula 1, observed the strongest physiologically active.At those R ₂With X be not in formula 1 compound of hydrogen, preferred configuration is expressed as L and D respectively on the carbon of indication.

Consider also that in field of the present invention other has the compound of formula 1, wherein R ₃Be alkoxyl group, cycloalkyloxy, aralkoxy or be selected from amino, dialkyl amido, the alkoxyl group that hydroxyl, alkoxyl group and halogen replace by one or more.

Formula 1 compound can be used the known any method preparation of those skilled in the art.These methods are included in and form peptide chain between the amino acid; owing to exist amino, carboxyl and other reactive group in the amino acid molecular; the above-mentioned group and/or make these group activations, particularly carboxyl so need protection is so that finish specified reaction or make this reaction optimizing.

In general, during synthesis type 1 compound, use two kinds of operational paths.First kind of following fragment of technology utilization

Second kind of technology comprises with suitable sour R ₁CO ₂H carries out acidylate to the peptide with following formula.

In the example of being given, to illustrate some blocking group and activating group especially here.Yet the those of skill in the art of an industry will find out, also can use other blocking group or activating group.Being chosen in of specific blocking group depends on to a great extent whether required reagent is easy to get; Influence to quilt " protection " compound dissolution degree; Remove the difficulty or ease of protecting group, and whether other group influenced by it, just depend on it selectivity or it whether be easy to remove.

For example, need or hope at least protection amino and/or carboxyl in many reactions.For the synthetic selected route of peptide may need to remove one or another or remove this two said blocking groups simultaneously, so that allow at regenerated amino or the enterprising single step reaction of carboxyl; That is used blocking group is reversible, and as a rule, can be removed independently of one another.In addition, be the selection of a given amino protecting group, depend on the effect of said amino in the entire reaction scheme.Using to have the different amino protecting groups of sloughing complexity, also is like this with regard to carboxy protective group.These groups are known in the prior art, and " polypeptide is synthetic " (" the Peptide Synt-hesis ") that shown at Bodansky etc., second edition, John Wiley ﹠amp; Sons, N.Y.(1976); " protecting group of using in the organic synthesis " that Greene showed " Protective Groups in Drganic Synthesis ") John Wiley d Sons, N.Y.(1981); " protecting group in the organic chemistry " that Mcomie showed (" Protective Groups in Organic Chemistry ") Plenum Press, N.Y.(1973); And " comprehensive organic chemistry; the synthetic and reaction of organic compound " (" Comprehensive Organic Chemis-try; TheSynthesis and Reactions of; Organic Compounds " that Sheppard showed,) Pergaman Press, N.Y.(1979), publish by E.Haslam, 23.6 section was carried out summary in 321～339 pages of books such as grade.

Habitual amino and carboxy protective group are known for industry those skilled in the art.Representational amido protecting group is as follows, is not limited to these certainly: carbobenzoxy-(Cbz) for example; Replace and unsubstituted aralkyl and benzyl, trityl, diphenyl-methyl and 4-nitrobenzyl; Benzylidene; Arylthio such as thiophenyl, nitre thiophenyl and trichlorobenzene sulfenyl; Phosphoryl derivative such as solutions of dimethyl phosphoryl base and O, O-dibenzyl phosphoryl; The trialkylsilkl derivative is as trimethyl silyl; With other as at United States Patent (USP) 4,322,341 described those groups.This patent is also here introduced with for referencial use.Preferred amino protecting group is a carbobenzoxy-(Cbz).The operation that replaces above-mentioned group on specified amino is known.General they comprise with Carbobenzoxy Chloride (benzyl chloroformate) in a kind of inert reaction solvent in the presence of alkali (acceptor of acid), suitable aminocompound is carried out acidylate.Used inert solvent has water, methylene dichloride, tetrahydrofuran (THF), and when with water as solvent, used alkali is sodium hydroxide or potassium hydroxide; When using organic solvent, then used alkali is tertiary amine, as contains the trialkyl amines and the pyridine of 1 to 4 carbon atom.When a kind of water solvent of use system, the pH of reaction is maintained at about 8～10, is preferably in pH9.In addition, when its amino needed protected reactant to contain basic group, itself just can be used as acid acceptor.

Acidylate operation with standard can be acyl group R ₁CO introduces in the peptide, as in an inert reaction solvent with suitable acyl chlorides or acylbromide and said reactive polypeptide.Advantageous conditions is the condition of non-water, comprising a kind of suitable alkali of adding, and for example a kind of organic bases, preferably a kind of tertiary amine is as triethylamine, N-methylmorpholine or pyridine.Best solvent is a methylene dichloride.

Representational carboxy protective group is various ester classes such as silyl ester, comprises trialkylsilkl ester, three halo silyl esters and haloalkyl silyl ester; Some hydrocarbyl carbonate, as contain benzyl ester, benzhydryl ester and the trityl ester of alkyl ester, the particularly tertiary butyl, benzyl and the replacement of 1 to 4 carbon atom; Benzoyl ester and phthalimido methyl esters; Some substituted hydrocarbon radical ester such as chloromethyl ester, 2,2,2-trichloro ethyl ester, cyanogen methyl esters; Tetrahydrofuran ester; The methoxy methyl esters; First sulphur methyl esters; Protected carbazyl is as-CONH-NHR ⁰, R wherein ⁰It is aforesaid an amido protecting group, particularly carbobenzoxy-(Cbz); And other is as at United States Patent (USP) 4,322, the group described in 341, and this patent is also listed in as a reference at this.Best carboxyl-protecting group is a tertbutyloxycarbonyl.

By the operation known to the industry technician, can change into unprotected amino and carboxyl to protected amino and carboxyl.Benzyl-for carboxyl (as the part of protected card azoles base) best protection base; can pass through palladium; particularly palladium carbon catalyst carries out catalytic hydrogenation and removes; in addition; also can be used on the trifluoromethanesulfonic acid in the trifluoroacetic acid; having in the presence of the methyl-phenoxide that suppresses alkylating, remove above-mentioned blocking group.By handling, can remove tertbutyloxycarbonyl at an easy rate with the saturated De diox of hydrogenchloride.

Quickening specified reaction by activated carboxyl is a kind of known method for those skilled in the art.Useful especially in the described here reaction sequence is the acid anhydride class, especially the cyclic anhydride class; And Acibenzolar class such as those Acibenzolar of deriving out from N-hydroxyphthalimide and N-maloyl imines, this two class all is used for the synthetic of peptide.

Activatory N-maloyl imines ester can quicken the follow-up reaction on said Acibenzolar group.As those skilled in the art understood, also available other activating group.A You Xingqu group is a N-hydroxyphthalimide base especially, and the usage of this group is the same with N-maloyl imido grpup.In these two examples, all be to form Acibenzolar with a kind of coupler that dewaters.Representational coupler has: 1-cyclohexyl-3-(2-morpholine ethyl)-carbodiimide-right-tosylate, dicyclohexyl carbodiimide, N, N ¹-carbonyl dimidazoles, the N-(3-dimethyl aminopropyl)-N ¹-ethyl-carbodiimide hydrochloride, ethoxy acetylene, diphenyl ketene and N-ethyl-5-isoxazole alkene-3 '-sulfonate.Use the reaction conditions of this coupler to be described in detail in the literature.General they comprise and use a kind of inert reaction solvent and from the temperature range of normal temperature to 100 ℃.Be good with above-mentioned carbodiimide reagent,, and obtain required ester with satisfied yield because they can react in room temperature.

After finishing the coupled reaction that causes final product, available suitable technology discussed above is removed various blocking groups, and formula 1 compound separation is come out.

As the alkali or the salt of pharmaceutical formula 1 compound, wherein R ₃Be hydroxyl, R ₄Or R ₅Be hydrogen, solution that can be by wushu 1 compound, the aqueous solution preferably, the alkali of having said with a kind of front reacts and makes by the ratio of stoichiometric requirement.The salt that forms is isolated with evaporation or sedimentary method.

Product of the present invention can be used as and comprises human mammiferous medicine, for because various cause of diseases are microbial, particularly clinical the and treatment of the disease that causes of Gram-negative bacteria is handled.

Comprise that product of the present invention also can be used as immunostimulant when human Mammals had the risk of infection of enhancement owing to the existing or clinical immunosuppression that causes.

The C that employing comes from Charles River Breeding laboratory ₃The H/HeN male mice, test method is as follows.Make mouse adapt to water and soil 5 days before the use earlier, use the various thinners (100,10,1 and 0.1 milligram/kilogram) or the placebo (pyrogen-free saline at) of test compound then, its used capacity is 0.2 milliliter, or makes subcutaneous (SC) or oral (PO) treats.The treatment system depends on the microorganism of used infection: healthy mice is done the immunity test of pneumobacillus and was treated in preceding 24 and 0 hours; The mouse of immunity infringement is cooked the immunity test of intestinal bacteria or streptococcus aureus and treated in preceding 3,2 and 1 days.The immunity test of pneumobacillus is to adopt in the muscle of hip injection, if peritoneal injection is then used in the immunity of intestinal bacteria and streptococcus aureus test, is used as the immunity test with 0.2 milliliter capacity.Pneumobacillus is then write down mortality ratio after 7 days in this way; Other two kinds of bacterium were then write down mortality ratio after 3 days in this way.

The preparation method of culture: pneumobacillus, intestinal bacteria and streptococcus aureus, containing on agar (BHI) substratum of brain and heart leach liquor, carry out streak culture purity with check frozen blood sample.The plate that three kinds of bacterium colonies picked up from 18 hours is cultivated, and puts it in 9 milliliters the BHI meat soup.Make broth culture under 37 ℃, growth is 2 hours in a rotation shaking machine, rules on the surface of several BHI agar slants with 0.2 milliliter then.After breeding 18 hours under 37 ℃,, adjust the density of culture and suitably dilution, make healthy mice obtain the standard of 90% lethal quantity (LD90) immunity test with Spect-ronic 20 with BHI meat soup washing inclined-plane.

When as human anti-infection agent or immunostimulant, The compounds of this invention is generally with prescription form oral administration, subcutaneous, muscle, intravenously, endoperitoneal administration at an easy rate.This prescription comprises the pharmacy routine.For example they can be to contain vehicle such as starch, lactose, tablet, pill, powder or the particle administration of the potter's clay of a definite form etc.They also can be taken with capsule (mixing to measure together or suitable vehicle) form.Also suspensoid that can be oral, solution, emulsion, syrup and elixir are taken, and wherein can contain spices and tinting material.For oral, the therapeutical agent that the present invention does contains about 50～500 milligrams tablet or capsule and in most of the cases is suitable for.

The doctor will determine for the optimum dosage of individual patients, and this dosage changes with the reaction and the route of administration of age, body weight and especial patient.Best oral dosage scope, (single dose or divided dose) are about 1.0 to about 300 milligrams/kilogram/days.Suitable parenteral dosage is about 1.0 to 100 milligrams/kilogram/days; About 1.0 to 20 milligrams/kilogram/days of preferred range.

The present invention also provides the formula of medicine that comprises unit dosage, and this prescription compound as described herein to using, that be used for disclosed purposes is valuable.Dosage form can single dose or multiple doses give, such as the aforementioned so that obtain to the effective per daily dose of special purpose.

The invention provides following embodiment for further illustrating, for for simplicity, following shortenings is used for the peak of nuclear magnetic resonance spectrum: S is unimodal; D refers to doublet; T refers to triplet; Q refers to quartet; M refers to multiplet.Mole (mole) and mmole (millimole) are abbreviated as m and mm respectively.

Embodiment 1

N-oenanthyl-D-γ-Gu Anxianji-glycyl-D-L-Ala

(R ₁=CH ₃(CH ₂) ₅; R ₂=H; And R ₃=NH H(CH ₃) CO ₂H)

1A.N-oenanthyl-D ₁-γ-Gu Anxianji (α-benzyl ester)-glycine

To be dissolved in the solution of triethylamine in 10 ml waters that 5.0 in 100 milliliters of dioxs grams (11.2 mmole) N-oenanthyl-D-γ-Gu Anxianji (α-benzyl ester)-maloyl imines ester is added to the glycine of 897 milligrams (13.0 mmoles) and 1.3 grams (13.0 mmole), the reaction mixture that obtains was stirring at room 80 hours.This solution is poured in 300 milliliters of ethyl acetate, and isolating organic phase is washed with 10% hydrochloric acid, water and salt brine solution in succession.Separate organic phase, use dried over mgso, be evaporated to driedly, resistates grinds with diethyl ether, and filters under nitrogen, 3.43 restrain (yield 74%).

1B.N-oenanthyl-D-gamma-glutamyl-glycyl-D-L-Ala

1-cyclohexyl-3-(2-morpholine ethyl with 3.03 grams (7.17 mmole)) carbodiimide N-first-P-TOLUENE SULFO ACID 99's salt is added to the D-L-Ala benzyl ester of N-oenanthyl-D-γ-Gu Anxianji (α-benzyl ester)-glycine, 1.75 grams (5 mmole) of 2.0 grams (4.78 mmole) right-triethylamine of tosylate, 506 milligrams (5 mmoles) and the I-hydroxybenzotriazole of 675 milligrams (5 mmoles) is in the solution of 100 milliliters of tetrahydrofuran (THF)s, and reaction mixture was stirring at room 18 hours.Reaction mixture poured in 300 milliliters the ethyl acetate, separate organic phase, and wash in succession with 10% hydrochloric acid, water, saturated sodium bicarbonate solution and salt solution.Organic phase is separated, and with dried over mgso and concentrating under reduced pressure, resistates grinds with ethyl acetate, filters under nitrogen, gets 2.7 grams.In 75 ml methanol, is 3.52 kilograms per centimeter at initial pressure with 2 gram solids and 400 milligram of 10% palladium carbon catalyst ²Hydrogen pressure under jolting 4 hours.Catalyzer is filtered out, and filtrate decompression concentrates, and the water-soluble and lyophilize of resistates gets 1.23 gram (yield 90%) white solid purpose products.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

4.35-4.2（m，2H），3.83（s，2H），2.35（t，J＝7Hz，2H），2.17（t，J＝7Hz，2H），2.1-1.8（m，2H），1.55-1.45（m，2H），1.3（d，J＝6Hz，3H），1.17（bs，6H）and????0.75（bs，3H）ppm.

Embodiment 2

N-oenanthyl-D-gamma-glutamyl-glycine (R ₁=CH ₃(CH ₂) ₅; R ₂=H; And R ₃=OH)

Handle N-oenanthyl-D-gamma-glutamyl (α-benzyl ester)-solution of glycine in 50 ml methanol that contains 1.0 grams with 100 milligram of 10% palladium carbon catalyst, in 3.52 kilograms per centimeter ²Hydrogen pressure under jolting 3 hours.With the catalyzer elimination, filtrate decompression concentrates.Resistates is dissolved in hot water, concentrating under reduced pressure.Resistates is dissolved in water again, and lyophilize, the purpose product of 630 milligrams of (yield 83%) white solids.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

4.37-4.25（m，1H），3.9（s，2H），2.35（t，J＝7Hz，2H），2.18（t，J＝6Hz，2H），2.4-1.8（m，2H），1.6-1.4（m，2H），1.8（bs，6H）and????0.7（bt，3H）ppm.

Embodiment 3

N-oenanthyl-D-gamma-glutamyl-glycyl-glycine

(R ₁=CH ₃(CH ₂) ₅; R ₂=H; And R ₃=NHCH ₂CO ₂H)

3A.N-oenanthyl-D-gamma-glutamyl (α-benzyl ester)-glycine maloyl imines ester

The dicyclohexyl carbodiimide of 7.0 grams (34 mmole) is added to N-oenanthyl-D-γ-Gu Anxianji (α-benzyl ester) glycine of 13.0 grams (31 mmole) and the N-maloyl imines (0 ℃) in the cold soln of 400 milliliters tetrahydrofuran (THF) of 3.91 grams (34 mmole); mixture stirred stirring at room 18 hours 1 hour at 0 ℃.Solid filtering is fallen, concentrates under the filtrate decompression, resistates with ether grind under nitrogen, filter the purpose intermediate of 15.4 grams (98%).

3B.N-oenanthyl-D-γ-Gu Anxianji-glycyl-glycine

The triethylamine that is dissolved in 446 milligrams of (5.95 mmole) glycine in 10 ml waters and 0.55 milliliter (3.9 mmole) is joined in 100 milliliters of dioxane solutions that contain 2.0 gram (3.97 mmole) N-oenanthyl-D-γ-Gu Anxianji (α-benzyl ester)-glycine maloyl imines esters, and the gained reaction mixture was stirring at room 18 hours.Solution is poured in 100 milliliters of ethyl acetate, with 2.5% hydrochloric acid, water and salt brine solution washing organic layer.Separate organic layer, with dried over mgso and be concentrated into dried.Resistates grinds with ethyl acetate, under nitrogen, filter 1.7 the gram white solids.With the solids of 1.5 grams in containing 200 milligrams of 75 ml methanol that are stated from 10% palladium hydroxide on the carbon, in 3.52 kilograms per centimeter ²Hydrogen pressure under jolting 3 hours.The elimination catalyzer, filtrate decompression concentrates, the water-soluble and dry purpose product that gets 1.12 grams (yield 90%) of freeze-drying of resistates.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.2-8.0（m，3H），4.19（m，1H），4.8-4.6（m，4H），2.25（t，J＝7Hz，2H），2.1（t，J＝6Hz，2H），2.05-1.7（m，2H），1.5（m，2H），1.25（bs，6H）and????0.85（t，J＝6Hz，3H）ppm.

Embodiment 4

N-oenanthyl-D-gamma-glutamyl-glycyl-D-Serine

D

(R ₁=CH ₃(CH ₂) ₅; R ₂=H; And R ₃=-NHCH(CH ₂OH) CO ₂H)

Starting raw material with 2.0 the gram (3.98 mmole) N-oenanthyl-D-γ-Gu Anxianji (α benzyl ester)-glycine maloyl imines ester, the O-benzyl-D-Serine of 780 milligrams (4.02 mmoles) and the triethylamine of 0.556 milliliter (4.02 mmole); and the operation of pressing embodiment 3B; separate 902 milligrams of purpose products (yield 76%), 130～132 ℃ of fusing points.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.36-7.94（m，3H），4.46-4.28（m，1H），4.28-4.08（m，1H），3.94-3.50（m，4H），2.25（t，J＝9Hz，2H），2.17（t，J＝9Hz，1H），2.10-1.04（m，14H）and????0.9（t，J＝6Hz，3H）ppm.

Embodiment 5

N-oenanthyl-D-gamma-glutamyl-glycyl-D-butyrine

(R ₁=CH ₃(CH ₂) ₅-; R ₂=H; And R ₃=-NH H(CH ₂CH ₃) CO ₂H

Repeat the operation of embodiment 3B, but starting raw material with 2.0 the gram (3.98 mmole) N-oenanthyl-D-gamma-glutamyl (α benzyl ester)-glycyl maloyl imines ester, the D-butyrine of 400 milligrams (4.02 mmoles) and the triethylamine of 0.556 milliliter (4.02 mmole), get the purpose product of 632 milligrams (yields 57%), 140～141 ℃ of fusing points.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.16-8.04（m，3H），4.22-4.08（m，2H），3.84-3.58（m，2H），2.2（t，J＝9Hz，2H），2.12（t，J＝9Hz，2H），2.04-1.0（m，15H）and????0.85（t，J＝6Hz，6H）ppm.

Embodiment 6

N-oenanthyl-D-gamma-glutamyl-glycyl-3-alanine

(R ₁=CH ₃(CH ₂) ₅-; R ₂=H; And R ₃=-NH(CH ₂) ₂CO ₂H)

Operation according to embodiment 3B, starting raw material with 1.5 the gram (3.0 mmole) N-oenanthyl-D-gamma-glutamyl (α benzyl ester)-glycine maloyl imines ester, the 3-alanine of 350 milligrams (3.9 mmoles) and the triethylamine of 0.55 milliliter (3.9 mmole), obtain 500 milligrams of (yield 43%) purpose products, 135～138 ℃ of fusing points.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.19-8.02（m，2H），7.98-7.87（t，J＝5Hz，1H），4.25-4.1（m，2H），3.8-3.49（m，2H），3.44-3.1（m，2H），2.4（t，J＝6Hz，2H），2.22（t，J＝7Hz，2H），2.14（t，J＝7Hz，2H），2.1-1.67（m，2H），1.6-1.17（m，8H）and????0.88（t，J＝6Hz，3H）ppm.

Embodiment 7

N-oenanthyl-D-gamma-glutamyl-glycyl-4-aminobutyric acid

(R ₁=CH ₃(CH ₂) ₅-; R ₂=H; And R ₃=-NH(CH ₂) ₃CO ₂H)

Repeat the operation of embodiment 6, the 4-aminobutyric acid replacement 3-alanine with 410 milligrams (4.0 mmoles) gets 600 milligrams of (yield 50%) purpose products, 140～142 ℃ of fusing points.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.18-8.03（m，2H），7.88（bt，J＝4Hz，1H），4.17-4.09（m，2H），3.81-3.48（m，2H），2.32-2.08（m，6H），2.08-1.08（m，12H）and????0.88（t，J＝6Hz，3H）ppm.

Embodiment 8

N-oenanthyl-D-gamma-glutamyl-glycyl-5-aminovaleric acid

(R ₁=CH ₃(CH ₂) ₅-; R ₂=H; And R ₃=-NH(CH ₂) ₄CO ₂H)

5-aminovaleric acid with 470 milligrams (4.0 mmoles) replaces the 3-alanine, and the method for pressing embodiment 6 obtains 520 milligrams of (yield 42%) purpose products, 122～124 ℃ of fusing points.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.25-7.94（m，2H），7.85（t，J＝5Hz，1H），4.25-4.1（m，2H），3.82-3.46（m，2H），3.24-2.9（m，2H），2.21-2.08（m，6H），2.08-1.2（m，14H）and????0.88（t，J＝6Hz，3H）ppm.

Embodiment 9

N-oenanthyl-D-gamma-glutamyl-glycyl-6-aminocaprolc acid

(R ₁=CH ₃(CH ₂) ₅-; R ₂=H; And R ₃=-NH(CH ₂) ₅CO ₂H)

Repeat the step of embodiment 6 again,, get the purpose product of 520 milligrams of (yield 40%) white foams with the 6-aminocaprolc acid replacement 3-alanine of 530 milligrams (4.0 mmoles).

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.28-7.9（m，2H），7.82（bt，J＝4Hz，1H），4.27-4.1（m，2H），3.81-3.47（m，2H），3.15-2.90（m，2H），2.3-2.08（m，6H），2.08-1.18（m，16H）and????0.88（t，J＝6Hz，3H）ppm.

Embodiment 10

N-isovaleryl-D-gamma-glutamyl-glycyl-D-L-Ala

(R ₁=(CH ₃) ₂CHCH ₂-; R ₂=H; And R ₃=-N

CH(CH ₃) CO ₂H)

10A. glycyl-D-L-Ala benzyl ester hydrochloride

The dicyclohexyl carbodiimides of 12.3 grams (60 mmole) are added to 100 milliliters of D-L-Ala benzyl ester tosilate and 5.77(57 mmoles that contain 10 gram (57 mmole) N-t-butoxycarbonyl glycines, 20 grams (57 mmole)) in cold (0 ℃) solution of methylene dichloride of triethylamine, the gained reaction mixture is warmed to room temperature.After 18 hours, mixture is filtered, filtrate decompression concentrates, and resistates is dissolved in 200 milliliters the ethyl acetate, and organic layer washs in succession with 2.5% hydrochloric acid, water, saturated sodium bicarbonate and salt solution.Separate organic layer, with dried over mgso and concentrating under reduced pressure.In the oil that obtains, add 200 milliliters with the saturated De diox of hydrogenchloride.After 30 minutes, add 400 milliliters of ether, under nitrogen, filter, get 10.9 gram products (yield 70%).

10B.N-tertbutyloxycarbonyl-D-gamma-glutamyl (α benzyl ester) maloyl imines ester

The dicyclohexyl carbodiimides of 30.9 grams (15 mmole) are added in 1500 milliliters of methylene dichloride that contain 50 gram (143 mmole) N-tertbutyloxycarbonyl-D-gamma-glutamic acid α-benzyl esters and 17.3 gram (150 mmole) N-maloyl imines, and the gained reaction mixture was stirring at room 18 hours.With solid filtering, filtrate decompression concentrates.Resistates grinds with ether, and solid filters under nitrogen, gets 43.7 grams (yield 68%).

10C.D-gamma-glutamyl (α benzyl ester)-glycyl-D-L-Ala benzyl ester hydrochloride

To contain 4.3 gram (9.45 mmole) N-tertbutyloxycarbonyl-D-gamma-glutamyl (α benzyl ester) maloyl imines esters, 2.71 gram (9.92 mmole) glycyl-D-L-Ala benzyl ester hydrochlorides and the solution of 1.0 gram (9.92 mmole) triethylamines in 100 milliliters of methylene dichloride, at room temperature stirred 18 hours, then concentrating under reduced pressure.Resistates is dissolved in 200 milliliters of ethyl acetate, and solution washs in succession with 2.5% hydrochloric acid, water, 10% solution of potassium carbonate and salt brine solution.Separate organic phase, use dried over mgso, reduction vaporization.Resistates is handled and was stirred 2 hours with the saturated De diox of hydrogenchloride with 200 milliliters.That solution decompression is concentrated into is dried, resistates grinds with ether.Solid is filtered under nitrogen, get 3.41 grams (yield 73%).

10D.N-isovaleryl-D-gamma-glutamyl-glycyl-D-L-Ala

490 milligrams of (4.06 mmole) isoveryl chloride are added in 1.0 gram (2.03 mmole) D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-L-Ala benzyl ester hydrochlorides and the solution of 616 milligrams of (6.09 mmole) triethylamines in 50 milliliters of methylene dichloride; reaction mixture was stirring at room 80 hours, and pressure reducing and steaming methylene dichloride, resistates are dissolved in the ethyl acetate.With gained solution with 2.5% hydrochloric acid, water, 10% salt of wormwood, water, and salt solution wash in succession.Separate organic phase, use dried over mgso, concentrating under reduced pressure.Resistates grinds with ether, filters (910 milligrams) under nitrogen.700 milligrams are dissolved in 50 ml methanol.Add 200 milligrams of palladium hydroxides, this mixture is in 3.52 kilograms per centimeter ²Jolting is 3 hours under the hydrogen pressure.With the catalyzer elimination, the pressure reducing and steaming solvent.Water-soluble and the lyophilize of resistates, the purpose product of 364 milligrams (yields 65%).

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.25-8.05（m，3H），4.33-4.12（m，2H），3.72（d，J＝6Hz，2H），2.21（t，J＝8Hz，2H），1.88-1.68（m，1H），2.08-1.9（m，4H），1.28（d，J＝9Hz，3H）and????0.9（d，J＝7Hz，6H）ppm.

Embodiment 11

With suitable chloride of acid and D-gamma-glutamyl (α benzyl ester)-glycyl-D-L-Ala benzyl ester hydrochloride as starting raw material, and with the step of embodiment 10D, prepared following compounds:

R ₁℃ m.p.. nucleus magnetic resonance, ppm

CH ₃（CH ₂） ₈- ＞175 （DMSO-d ₆）8.17-8.22

（m，2H），4.35-4.13

（m，2H），3.83（d，J＝

6Hz，2H），2.22（t，

J＝6Hz，2H），2.15（t，

J＝6Hz，2H），2.08-

1.65（m，2H），1.45-

1.18(m, 17H) and

0.90（t，J＝7Hz，3H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

（CH ₃） ₂CH- ＞110（dec） （DMSO-d ₆）8.2-8.08

(decomposition) (m, 2H), 8.02(d, J=

9Hz，1H），4.3-4.1（m，

2H），3.72（d，J＝7Hz，

2H），2.53-2.40（m，

1H），2.22（t，J＝9Hz，

2H），2.10-1.7（m，

2H），1.28（d，J＝9Hz，

3H) and 1.08-0.9

（m，6H）

＞110（dec） （DMSO-d ₆）8.3-7.9

(decomposition) (m, 3H), 4.33-4.17

（m，2H），3.72（d，

J＝5Hz，2H），2.3-2.1

(m, 2H) and 2.10-1.0

（m，16H）

CH ₃（CH ₂） ₂- ＞110（dec） （DMSO-d ₆）8.25-8.05

(decomposition) (m, 3H), 4.35-4.22

（m，2H），3.72（d，J＝

6Hz，2H），2.22（t，

J＝8Hz，2H），2.12（t，

J＝8Hz，2H），2.08-

1.77（m，2H），1.55

（q，J＝8Hz，2H），1.28

(d, J=8Hz, 3H) and

0.88（t，J＝7Hz，3H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

CH ₃（CH ₂） ₄- ＞190（dec） （DMSO-d ₆）8.33-8.0

(decomposition) (m, 3H), 4.35-4.1

（m，2H），3.7（d，J＝

6Hz，2H），2.17（t，

J＝8Hz，2H），2.1（t，

J＝8Hz，2H），2.05-

1.63（m，2H），1.48

（t，J＝7Hz，2H），1.2-

1.1(m, 7H) with 0.85

（t，J＝7Hz，3H）

CH ₃（CH ₂） ₆- ＞180（dec） （D ₂O）4.42-4.28

(decomposition) (m, 2H), 3.91(s,

1H），2.4（t，J＝7Hz，

2H），2.27（t，J＝7Hz，

2H），2.22-1.94（m，

2H），1.65-1.55（m，

2H），1.39（d，J＝8Hz，

3H），1.34-1.17（m，

8H) and 0.73(m, 3H)

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.27-8.03

（m，3H），4.32-4.1

（m，2H），3.72（d，

J＝6Hz，2H），2.22

（t，J＝10Hz，2H），

2.27-1.68（m，6H），

1.42-1.0（m，10H）

And 0.94-0.8(m, 6H)

- （DMSO-d ₆）8.22-8.0

（m，3H），4.32-4.1

（m，2H），3.8-3.6

（m，2H），2.28-1.68

（m，6H），1.6-1.0

(m, 12H) and 0.94-

0.7（m，6H）。

（CH ₃CH ₂） ₂CH- - （DMSO-d ₆）8.29-7.97

（m，3H），4.33-4.1

（m，2H），3.81-3.59

（m，2H），2.32-1.65

（m，6H），1.65-1.17

(m, 8H) and 1.02-

0.68（m，6H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.3-8.0

（m，3H），4.32-4.1

（m，2H），3.85-3.62

（m，2H），2.21（t，

J＝8Hz，2H），2.02

（d，J＝8Hz，2H），

2.01-1.9（m，1H），

1.85-1.5（m，8H），

1.28（d，J＝8Hz，3H）

And 1.28-0.8(m, 5H)

（CH ₃CH ₂CH ₂） ₂CHCH ₂- （DMSO-d ₆）8.18-8.0

（m，3H），4.31-4.1

（m，2H），3.84-3.6

（m，2H），2.22（t，

J＝6Hz，2H），2.07

（d，J＝8Hz，2H），

2.03-1.7（m，3H），

1.4-1.15（m，11H）

And 0.87(t, J=6Hz,

6H）

（CH ₃CH ₂） ₂CHCH ₂- - （DMSO-d ₆）8.27-7.95

（m，3H），4.3-4.1（m，

2H），3.78-3.6（m，

2H），2.3-1.57（m，

8H），1.46-1.13（m，

8H) and 0.84(t, J=

8Hz，6H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.18-8.0

（m，3H），4.24-4.06

（m，2H），3.74-3.56

（m，2H），2.17（t，J＝

9Hz，2H），2.12-2.0

（m，1H），2.0-1.64

（m，4H），1.24（d，

J＝6Hz，7H），1.14-

0.98(m, 2H) and

0.81（d，J＝6Hz，6H）

- （DMSO-d ₆）8.2-8.04

（m，3H），4.26-4.08

（m，2H），3.76-3.6

（m，2H），2.28-1.64

（m，7H），1.4-0.96

(m, 7H) and 0.96-

0.74（m，6H）。

R ₁℃ m.p.. nucleus magnetic resonance, ppm

（CH ₃） ₂CH（CH ₂） ₃- - （DMSO-d ₆）8.24-7.95

（m，3H），4.3-4.08

（m，2H），3.81-3.59

（m，2H），2.21（t，

J＝6Hz，2H），2.11

（t，J＝8Hz，2H），

2.05-1.38（m，7H），

1.27（d，J＝8Hz，3H），

1.17-1.05（m，2H）

And 0.86(d, J=10Hz,

6H）

- （DMSO-d ₆）8.24-8.0

（m，3H），4.33-4.11

（m，2H），3.79-3.6

（m，2H），2.41-2.29

（m，1H），2.22（t，

J＝8Hz，2H），2.11-1.66

（m，2H），1.59-1.43

（m，1H），1.38-1.11

（m，11H），1.06-0.95

(m is 3H) with 0.87

（t，J＝6Hz，3H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.24-8.02

（m，3H），4.32-4.11

（m，2H），3.84-3.6

（m，2H），2.24（t，

J＝8Hz，2H），2.08

（d，J＝8Hz，2H），2.03-

1.63（m，4H），1.44-

1.11(m, 12H) and

0.97-0.71（m，6H）

（CH ₃） ₂CH（CH ₂） ₄- - （DMSO-d ₆）8.24-8.04

（m，3H），4.28-4.1

（m，2H），3.76-3.6

（m，2H），2.18（t，J＝

6Hz，2H），2.1（t，

J＝6Hz，2H），2.04-

1.86（m，1H），1.84-

1.66（m，1H），1.56-

1.38（m，3H），1.23

（d，J＝6Hz，3H），1.2-

1.06(m, 3H) and

0.82（d，J＝6Hz，6H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.23-7.98

（m，3H），4.3-4.13

（m，2H），3.81-3.61

（m，2H），2.22（t，

J＝8Hz，2H），2.18-

1.68（m，6H），1.45-

1.07(m, 12H) and

0.98-0.8（m，6H，

- （DMSO-d ₆）8.37-8.03

（m，3H），4.31-4.1

（m，2H），3.78-3.6

（m，2H），2.26（t，

J＝8Hz，2H），2.2-1.36

（m，7H），1.3（d，J＝

8Hz，5H），1.26-1.05

(m, 2H) and 1.05-

0.73（m，9H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.23-7.98

（m，3H），4.3-4.13

（m，2H），3.81-3.61

（m，2H），2.21（t，

J＝8Hz，2H），2.15-

2.0（m，2H），1.9

（t，J＝8Hz，2H），

1.85-1.52（m，3H），

1.4-1.22（m，3H），

1.22-0.94（m，3H）

And 0.94-0.80(m, 6H)

- （DMSO-d ₆）8.25-7.98

（m，3H），4.3-4.08

（m，2H），3.81-3.62

（m，2H），2.22（t，

J＝8Hz，2H），2.06

（d，J＝8Hz，2H），

2.02-1.89（m，1H），

1.87-1.65（m，2H），

1.41-1.06（m，11H）

And 0.98-0.7(m, 6H)

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.33-7.95

（m，3H），4.3-4.06

（m，2H），3.83-3.59

（m，2H），2.21（t，

J＝8Hz，2H），2.11（s，

3H），2.08-1.87（m，

1H），1.87-1.35（m，

8H），1.25（d，J＝8Hz，

3H) and 1.22-0.98

（m，3H）

（CH ₃CH ₂） ₂CH（CH ₂） ₂- - （DMSO-d ₆）8.21-8.0

（m，3H），4.32-4.1

（m，2H），3.83-3.6

（m，2H），2.21（t，

J＝8Hz，2H），2.11

（t，J＝8Hz，2H），

2.05-1.89（m，1H），

1.87-1.67（m，1H），

1.57-1.38（m，3H），

1.38-1.08（m，9H）

And 0.83(t, J=6Hz,

6H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.24-7.97

（m，3H），4.33-4.1

（m，2H），3.86-3.59

（m，2H），2.35-2.08

（m，4H），2.08-1.9

（m，1H），1.89-1.67

（m，1H），1.63-1.46

（m，1H），1.46-1.02

(m, 11H) and 0.98-

0.73（m，6H）

- （DMSO-d ₆）8.2-7.94

（m，3H），4.26-4.06

（m，2H），3.76-3.56

（m，2H），2.16（t，

J＝6Hz，2H），2.1-1.84

（m，3H），1.84-1.6

（m，2H），1.24（d，J＝

6Hz，9H），1.12-0.92

(m, 3H) and 0.92-

0.64（m，9H）

R ₁℃ m.p.. nucleus magnetic resonance, ppm

- （DMSO-d ₆）8.23-8.0

（m，3H），4.32-4.06

（m，2H），3.72（d，

J＝8Hz，2H），2.22

（t，J＝10Hz，2H），

2.16-1.7（m，6H），

1.42-1.08（m，14H）

And 0.92-7.0(m, 6H)

- （DMSO-d ₆）8.2-8.0

（m，3H），4.24-4.16

（m，2H），3.74-3.60

（m，2H），2.18（t，

J＝7，2H），2.02（d，

J＝7，2H），2.02-1.6

（m，3H），1.26（d，

J＝6，3H），1.26-1.08

(m, 8H) and .92-.74

（m，6H）

Embodiment 12

N-(3-(S)-the methyl oenanthyl)-D-gamma-glutamyl-L-alanyl-D-L-Ala (R ₁=(S) CH ₃(CH ₂) ₃CH(CH ₃) CH ₂-; R ₂=CH ₃;

R ₃＝

（CH ₃）CO ₂H）

12A.N-tertbutyloxycarbonyl-L-alanyl-D-L-Ala benzyl ester

In the solution of 25.0 gram (0.121 mole) dicyclohexyl carbodiimides that will be in 100 milliliters of methylene dichloride are added drop-wise to the D-L-Ala benzyl ester of 23.0 gram (0.121 mole) N-tert-butoxycarbonyl-l-alanines in the methylene dichloride of 400 milliliters cold (0 ℃), 42.6 grams (0.121 mole) right-tosylate and 17 milliliters of (0.121 mole) triethylamines, in stirred overnight at room temperature, the elimination solid is condensed into oil with filtrate.Resistates is dissolved in after 400 milliliters the ethyl acetate with 1% hydrochloric acid soln, 10% solution of potassium carbonate, and water and salt solution wash in succession.Separate organic phase, use dried over mgso, and be condensed into oil.Resistates grinds with diethyl ether, the gained solid under nitrogen, filter 16.0 grams, 12.7 gram purpose product crystallizations from filtrate in addition.

12B.L-alanyl-D-L-Ala benzyl ester hydrochloride

Be dissolved at 28.7 gram N-tertbutyloxycarbonyl-L-alanyl-D-third ammonia benzyl esters and add 150 milliliters in the soup compound that 150 milliliters of dioxs form with the saturated De diox of hydrochloric acid.

Mixture was stirring at room 4 hours.Removal of solvent under reduced pressure, resistates grinds with ether.Filter the gained solid, again it is dissolved in the methylene dichloride, this solution concentration to about 150 milliliters, is added ether, descended filter solid, get 22.0 grams at nitrogen.

12C.N-uncle-butoxy carbonyl-D-gamma-glutamyl (α benzyl ester)-L-alanyl-D-L-Ala benzyl ester

In the soup compound of 5 gram (9.64 mmole) N-tert-butoxycarbonyl-D-gamma-glutamyl amine α benzyl ester dicyclohexyl amines in being chilled to 100 milliliters of methylene dichloride of 0 ℃ and the L-alanyl-D-L-Ala benzyl ester hydrochloride of 2.76 grams (9.64 mmole), be added in the dicyclohexyl carbodiimide of 2.0 grams (9.64 mmole) in 20 milliliters of the same solvents.In stirred overnight at room temperature, the elimination solid, filtrate decompression concentrates.Resistates is with ethyl acetate 150 milliliters of processing, the elimination solid, filtrate is with 1% hydrochloric acid, 10% solution of potassium carbonate, water and salt solution wash in succession, the organic phase dried over sodium sulfate, concentrate white solid, with the ether grinding, filter the 4.1 purpose products that restrain.

12D.D-gamma-glutamyl (α benzyl ester)-L-alanyl-D-L-Ala benzyl ester hydrochloride

In 100 milliliters of soup compounies that are added in 4.1 in 50 milliliters of dioxs grams (7.21 mmole) uncle N--butoxy carbonyl-D-gamma-glutamyl (α benzyl ester)-L-alanyl-D-L-Ala benzyl ester with the saturated De diox of hydrogenchloride, reaction mixture was stirring at room 3 hours, removal of solvent under reduced pressure, resistates grinds with diethyl ether, gets 3.5 grams.

12E.N-(3-(S)-methyl oenanthyl)-D-gamma-glutamyl (α benzyl ester)-L-alanyl-D-L-Ala benzyl ester

3-(S with 390 milligrams (2.37 mmoles))-the methyl oenanthyl chloro is added in the triethylamine of 1.0 gram D-gamma-glutamyl (α benzyl ester)-L-alanyl-D-L-Ala benzyl esters of (1.98 mmole) in 50 milliliters of methylene dichloride and 0.833 milliliter (5.93 mmole).Reaction mixture stirred 45 minutes under nitrogen, reaction mixture was poured in 150 milliliters the ethyl acetate, and organic phase is washed in succession with 10% hydrochloric acid, 10% solution of potassium carbonate, water and salt solution.Organic phase dried over sodium sulfate, and be concentrated into driedly, resistates is developed with ether, under nitrogen, filter, 900 milligrams.

12F.N-(3-(S)-methyl oenanthyl)-D-γ-Gu Anxianji-L-alanyl-D-L-Ala

With 200 milligrams of palladium hydroxide and 900 milligrams of N-(3-(S in 50 ml methanol that are stated from carbon)-the methyl oenanthyl)-mixture of D-gamma-glutamyl (α benzyl ester)-L-alanyl-D-L-Ala benzyl ester is in 3.52 kilograms per centimeter ²Hydrogen pressure jolting 1 hour.The elimination catalyzer, the pressure reducing and steaming solvent.In resistates, add water and reduce pressure and remove, get 492 milligrams white solid product, 165～168 ℃ of fusing points.

Nuclear magnetic resonance spectrum (DMSO-d ₆) show following absorption:

8.21-7.98（m，3H），4.41-4.1（m，3H），2.3-2.06（m，4H），2.06-1.56（m，6H），1.43-1.02（m，11H）and????1.02-0.73（m，6H）ppm.

Embodiment 13

N-(3-(S, R)-ethyl hexanoyl)-D-gamma-glutamyl (α-n-butyl) glycyl-D-L-Ala (R ₁=CH ₃(CH ₂) ₂CH(C ₂H ₅) CH ₂-;

D

R ₂＝H;R ₃＝-

（CH ₃）CO ₂H;R ₄＝n-C ₄H ₉

13A.N-tertbutyloxycarbonyl-D-gamma-glutamyl amine (α n-butyl) dicyclohexyl amine salt

39.5 gram (0.172 mole) N-tertbutyloxycarbonyl-D-L-glutamic acid anhydride solutions that will be in 75 milliliters of dry tetrahydrofuran are in two hours, in 0 ℃ of propyl carbinol and the solution of 34.3 milliliters of (0.172 mole) dicyclohexylamine in 300 milliliters of ether that is added drop-wise to by 47 milliliters (0.516 moles), be reflected at 0 ℃ of stirring and carried out 3 hours and in water tank, place and spend the night.The elimination solid is mixed into pulpous state and filtration in ethanol, get 43.3 grams.

13B.D-gamma-glutamyl (α n-butyl)-glycyl-D-L-Ala benzyl

Ester hydrochloride

Product 10 grams (0.021 mole) of embodiment 13A and the glycyl-D-L-Ala benzyl ester hydrochloride of 6.7 grams (0.024 mole) are mixed into pulpous state with 200 milliliters of methylene dichloride, and are as cold as 0 ℃ under nitrogen.Add dicyclohexyl carbodiimide (0.021 mole of 4.25 gram), mixture is warmed to ambient temperature overnight.Remove by filter the urea by product, the solvent decompression is removed, resistates is handled and is filtered with ethyl acetate, and filtrate is water, 2.5% hydrochloric acid, water, 10% salt of wormwood and salt water washing successively.With organic phase dried over mgso, removal of solvent under reduced pressure.Resistates is dissolved in 300 milliliters with in the saturated De diox of hydrogenchloride., remove and desolvate after 4 hours in stirring at room, resistates gets 7.4 grams with ethyl acetate-hexane (1: 1) development and filtration.

13C.N-(3-(S, R)-thylhexoic acid)-D-glutamy (α n-butyl) glycyl-D-L-Ala

At the product that is dissolved in 50 milliliters of embodiment 13B in the methylene dichloride (1.0 grams, 2.35 mmole) and in the triethylamine of 0.99 milliliter (7.05 mmole), add 460 milligrams of (2.83 mmole) 3-(S, R) ethyl hexanoyl chloro, this reaction mixture stir under nitrogen and spend the night, removal of solvent under reduced pressure, resistates are dissolved in the ethyl acetate.Organic phase is used 10% hydrochloric acid, water, 10% salt of wormwood and salt water washing successively.Separate organic phase, use dried over mgso, removal of solvent under reduced pressure, resistates is dissolved in 10 ml methanol, and with 170 milligrams of 10% palladium hydroxides that are stated from the carbon be 3.52 kilograms per centimeter at initial pressure ²Hydrogen pressure under jolting 1.5 hours.With the catalyzer elimination that consumes, the solvent decompression is removed, and gets 100 milligrams.

Nucleus magnetic resonance (DMSO-d ₆):

NMR（DMSO-d ₆）：8.18（d，J＝6，1H），8.10（d，J＝6，1H），8.02（t，J＝5，1H），4.28-4.10（m，2H），4.00（t，J＝6，2H），3.78-3.56（m，2H），2.18（t，J＝6，2H），2.02（d，J＝6，2H），2.00-1.60（m，3H），1.58-1.42（m，2H），1.28-1.08（m，8H），1.24（d，J＝6，3H），0.92-0.76（m，9H）。

Embodiment 14

With the general operation method of embodiment 13, begin to have prepared following compounds from the reactant of necessity:

R ₁R ₄Nucleus magnetic resonance

3-methyl oenanthyl methyl N MR(DMSO-d ₆): 8.23(d,

J＝6，1H），8.15（d，J＝6，

1H），8.09（t，J＝6，1H），

4.28-4.14（m，2H）;3.72

（d，J＝6，2H），3.61（s，

3H），2.23（t，J＝7，2H），

2.16-1.70（m，6H），1.34-

1.04（m，8H），1.25（d，

J＝7，3H），0.92-0.76（m，

6H）.

R ₁R ₄Nucleus magnetic resonance

3-ethyl oenanthyl methyl N MR(DMSO-d ₆): 8.19(d,

J＝6，1H），8.11（d，J＝6，

1H），8.03（t，J＝6，1H），

4.24-4.10（m，2H），3.74-

3.62（m，2H），3.57（s，

3H），2.18（t，J＝9，2H），

2.01（d，J＝6，2H），1.97-

1.60（m，3H），1.32-1.10

（m，8H），1.23（d，J＝7，

3H），0.90-0.72（m，6H）

3-methyl oenanthyl ethyl n MR(DMSO-d ₆): 8.24-8.02

（m，3H），4.26-3.96（m，

2H），4.04（q，J＝9，2H），

3.76-3.56（m，2H），2.17

（t，J＝7，2H），2.12-1.63

（m，6H）;1.74-0.94（m，

6H），1.23（d，J＝5，3H），

1.13（t，J＝9，3H），0.88-

0.72（m，6H）

R ₁R ₄Nucleus magnetic resonance

3-ethyl oenanthyl ethyl n MR(DMSO-d ₆): 8.16(d,

J＝6，1H），8.09（d，J＝6，

1H），8.02（t，J＝6，1H），

4.22-4.08（m，2H），4.02

（q，J＝7，2H），3.74-3.54

（m，2H），2.16（t，J＝7，

2H），2.00（d，J＝6，2H），

1.96-1.58（m，3H），1.30-

1.08（m，8H），1.21（d，

J＝7，3H），1.13（t，J＝7，

3H），0.88-0.70（m，6H）

3-methyl oenanthyl isobutyl-NMR(DMSO-d ₆): 8.14(d,

J＝6，1H），8.08-7.98（m，

2H），4.20-4.04（m，2H），

3.75（d，J＝6，2H），3.68-

3.54（m，2H），2.14（t，

J＝6，2H），2.08-1.64（m，

6H），1.28-0.96（m，6H），

1.19（d，J＝7，3H），0.88-

0.70（m，12H）

R ₁R ₄Nucleus magnetic resonance

3-ethyl oenanthyl isobutyl-NMR(DMSO-d ₆): 8.21(d,

J＝6，1H），8.14-8.04（m，

2H），4.24-4.08（m，2H），

3.79（d，J＝6，2H），3.72-

3.58（m，2H），2.19（t，

J＝7，2H），2.03（d，J＝6，

2H），1.99-1.60（m，4H），

1.32-1.10（m，8H），1.23

（d，J＝6，3H），0.92-0.72

（m，12H）

3-ethyl hexanoyl isobutyl-NMR(DMSO-d ₆): 8.18(d,

J＝6，1H），8.10-8.00（m，

2H），4.26-4.08（m，2H），

3.79（d，J＝6，2H），3.72-

3.58（m，2H），2.18（t，

J＝6，2H），2.02（d，J＝6，

2H），1.98-1.62（m，4H），

1.34-1.08（m，6H），1.23

（d，J＝7，3H），0.96-0.72

（m，12H）

R ₁R ₄Nucleus magnetic resonance

3-ethyl hexanoyl methyl N MR(DMSO-d ₆): 8.21(d,

J＝7，1H），8.10（d，J＝7，

1H），8.05（t，J＝6，1H），

4.26-4.10（m，2H），3.76-

3.60（m，2H），3.59（s，

3H），2.18（t，J＝6，2H），

2.02（d，J＝6，2H），2.00-

1.60（m，3H），1.32-1.08

（m，7H），0.90-0.72（m，

6H）.

3-ethyl hexanoyl ethyl n MR(DMSO-d ₆): 8.22(d,

J＝7，1H），8.18-8.06（m，

2H），4.26-4.10（m，2H），

4.06（q，J＝5，2H），3.78-

3.58（m，2H），2.20（t，

J＝6，2H），2.04（d，J＝6，

2H），2.02-1.60（m，3H），

1.26-1.20（m，7H），1.18

（t，J＝5，3H），0.90-0.78

（m，6H）

R ₁R ₄Nucleus magnetic resonance

3-methyl oenanthyl butyl NMR(DMSO-d ₆): 8.20(d,

J＝7，1H），8.16-8.04（m，

2H），4.24-4.06（m，2H），

4.00（t，J＝6，2H），3.74-

3.56（m，2H），2.17（t，

J＝6，2H），2.12-1.60（m，

5H），1.58-1.40（m，2H），

1.36-1.00（m，8H），1.21

（d，J＝6，3H），0.90-0.74

（m，9H）

3-ethyl oenanthyl butyl NMR(DMSO-d ₆): 8.16(d,

J＝7，1H），8.11（d，J＝7，

1H），8.03（t，J＝5，1H），

4.26-4.09（m，2H），3.99

（t，J＝7，2H），3.79-3.58

（m，2H），2.17（t，J＝6，

2H），2.01（d，J＝6，2H），

2.00-1.60（m，3H），1.58-

1.42（m，2H），1.36-1.08

（m，10H），1.24（d，J＝5，

3H），0.92-0.72（m，9H）

Embodiment 15

N-(3-(R, S)-ethyl hexanoyl)-D-gamma-glutamyl-glycyl-D-alanine ethyl ester (R ₁=CH ₃(CH ₂) ₂CH(C ₂H ₅) CH ^- ₂; R ₂=H;

R ₃＝-

（CH ₃）CO ₂C ₂H ₅;R ₅＝H）

15A.D-gamma-glutamyl (α benzyl ester) glycyl-D-alanine ethyl ester hydrochloride

5.6 gram (0.0270 mole) dicyclohexyl carbodiimides are added in the slurries of glycyl-D-L-Ala ethyl ester hydrochloride of 14.8 gram (0.0285 mole) N-tertbutyloxycarbonyl-D-gamma-glutamic acid α benzyl ester dicyclohexyl amine salts in 200 milliliters of methylene dichloride and 6 grams (0.0285 mole).This mixture stirs under nitrogen and spends the night.Cross elimination urea, removal of solvent under reduced pressure.Resistates is handled with 300 milliliters of ethyl acetate, filters.Filtrate is successively with 2.5% hydrochloric acid, water, 10% solution of potassium carbonate and salt water washing.Separate organic phase, use dried over mgso, concentrating under reduced pressure.Irreducible oil is dissolved in 450 milliliters of saturated De dioxs of hydrogenchloride.Stirred this solution 2 hours, removal of solvent under reduced pressure.The development of resistates and ether and filter 11.2 grams.

15B.N(3-(R, S)-ethyl hexanoyl)-D-gamma-glutamyl-glycyl-D-alanine ethyl ester

With the 3-(R of 378 milligrams (2.33 mmoles), S)-ethyl hexanoyl chloro is added under nitrogen atmosphere in the triethylamine of the product (1.0 gram, 2.33 mmoles) of the embodiment 15A in 30 milliliters of methylene dichloride and 0.98 milliliter (6.98 mmole).After at room temperature stirring 1.5 hours, this mixture is poured in 100 milliliters of ethyl acetate, organic phase is successively with 10% solution of potassium carbonate and salt water washing.Separate organic phase, use dried over mgso, concentrating under reduced pressure.The white solid resistates is dissolved in 30 ml methanol, and in 3.25 kilograms per centimeter ²Reduce with 0.1 gram palladium hydroxide in the nitrogen atmosphere of initial pressure.Elimination catalyzer after 2 hours, filtrate is concentrated into dried, and resistates gets 275 milligrams with ether development and filtration.

NMR（DMSO-d ₆）8.26（d，J＝9，1H），8.14-8.02（m，2H），4.31-4.00（m，2H），4.06（q，J＝10，2H），3.78-3.60（m，2H），2.17（t，J＝8，2H），2.08-1.65（m，1H），2.03（d，J＝8，2H），1.82-1.53（m，3H），1.40-0.96（m，5H），1.23（d，J＝6，3H），1.14（t，J＝10，3H），0.90-0.64（m，6H）.

Embodiment 16

From suitable reagent, adopt the method for embodiment 15A～15B, prepared following compounds:

R ₁R ₅Nucleus magnetic resonance

3-methyl oenanthyl isobutyl-NMR(DMSO-d ₆): 8.24

（d，J＝6，1H），8.10-8.00

（m，2H），4.30-4.18（m，

1H），4.18-4.08（m，1H），

3.86-3.72（m，2H），

3.72-3.58（m，2H），

2.16（t，J＝6，2H），

2.12-1.64（m，6H），

1.52-1.00（m，6H），

1.27（d，J＝7，3H），

0.90-0.76（m，12H）

3-ethyl hexanoyl isobutyl-NMR(DMSO-d ₆): 8.23(d,

J＝6，1H），8.08-7.99（m，

2H），4.29-4.17（m，1H），

4.17-4.07（m，1H），

3.83-3.71（m，2H），

3.71-3.58（m，2H），2.15

（t，J＝7，2H），2.04-1.60

（m，4H），2.00（d，J＝6，

2H），1.31-1.09（m，6H），

1.25（d，J＝6，3H），

0.90-0.72（m，12H）

R ₁R ₅Nucleus magnetic resonance

3-ethyl oenanthyl isobutyl-NMR(DMSO-d ₆): 8.23(d,

J＝6，1H），8.08-7.98（m，

2H），4.29-4.18（m，1H），

4.18-4.07（m，1H），

3.86-3.72（m，2H），

3.70-3.57（m，2H），2.15

（t，J＝7，2H），2.04-1.59

（m，4H），2.00（d，J＝6，

2H），1.30-1.11（m，8H），

1.25（d，J＝6，3H），

0.89-0.70（m，12H）

3-methyl oenanthyl methylcyclohexyl NMR(DMSO-d ₆): 8.25(d,

J＝6，1H），8.13-8.00（m，

2H），4.32-4.20（m，1H），

4.20-4.08（m，1H），

3.91-3.76（m，2H），

3.76-3.59（m，2H），2.18

（t，J＝6，2H），2.13-1.48

（m，8H），1.36-1.01（m，

12H），1.27（d，J＝6，

3H），1.02-0.76（m，8H）

R ₁R ₅Nucleus magnetic resonance

3-ethyl hexanoyl methylcyclohexyl NMR(DMSO-d ₆): 8.25(d,

J＝6，1H），8.13-8.00（m，

2H），4.32-4.20（m，1H），

4.20-4.08（m，1H），

3.92-3.74（m，2H），

3.74-3.59（m，2H），2.18

（t，J＝6，2H），2.09-1.86

（m，1H），2.03（d，J＝6，

2H），1.82-1.43（m，8H），

1.36-1.01（m，9H），1.27

（d，J＝6，3H），1.01-0.70

（m，8H）

3-ethyl oenanthyl methylcyclohexyl NMR(DMSO-d ₆): 8.26(d,

J＝6，1H），8.12-8.02（m，

2H），4.31-4.19（m，1H），

4.19-4.08（m，1H），

3.93-3.72（m，2H），

3.72-3.58（m，2H），

2.18（t，J＝6，2H），

2.08-1.86（m，1H），2.03

（d，J＝6，2H），1.82-1.48

（m，8H），1.34-1.02（m，

11H），1.27（d，J＝6，

3H），1.00-0.74（m，8H）

R ₁R ₅Nucleus magnetic resonance

3-methyl oenanthyl ethyl n MR(DMSO-d ₆): 8.25(d,

J＝6，1H），8.12-8.00（m，

2H），4.28-3.96（m，2H），

4.03（q，J＝7，2H），

3.74-3.56（m，2H），2.16

（t，J＝9，2H），2.11-1.62

（m，6H），1.32-0.98（m，

6H），1.24（d，J＝7，3H），

1.14（t，J＝7，3H），

0.88-0.76（m，6H）

3-ethyl oenanthyl ethyl n MR(DMSO-d ₆): 8.28(d,

J＝6，1H），8.16-8.04（m，

2H），4.32-4.04（m，2H），

4.10（q，J＝6，2H），

3.78-3.64（m，2H），2.22

（t，J＝6，2H），2.11-1.92

（m，1H），2.07（d，J＝6，

2H），1.86-1.64（m，2H），

1.40-1.14（m，8H），1.30

（d，J＝6，3H），1.21（t，

（J＝6，3H），0.94-0.76

（m，6H）.

R ₁R ₅Nucleus magnetic resonance

3-ethyl oenanthyl butyl NMR(DMSO-d ₆): 8.27(d,

J＝8，1H），8.14-8.02（m，

2H），4.32-4.10（m，2H），

4.10-3.94（m，2H），

3.78-3.60（m，2H），2.18

（t，J＝6，2H），2.04（d，

J＝6，2H），2.04-1.62（m，

3H），1.60-1.46（m，2H），

1.38-1.10（m，12H），

1.27（d，J＝6，3H），

0.90-0.75（m，9H）

3-S-methyl oenanthyl butyl NMR(DMSO-d ₆): 8.30(d,

J＝8，1H），8.15-8.04（m，

2H），4.45-4.12（m，2H），

4.12-3.98（m，2H），

3.78-3.65（m，2H），2.22

（t，J＝7，2H），2.18-1.69

（m，7H），1.61-1.48（m，

2H），1.40-1.11（m，

11H），0.97-0.80（m，9H）

R ₁R ₅Nucleus magnetic resonance

3-S-ethyl oenanthyl butyl NMR(DMSO-d ₆): 8.22(d,

J＝7，1H），8.12-8.0（m，

2H），4.4-4.16（m，2H），

4.08-3.95（m，2H），

3.75-3.62（m，2H），2.18

（t，J＝6，2H），2.02（d，

J＝6，2H），2.04-1.62（m，

3H），1.60-1.46（m，2H），

1.38-1.1（m，15H），and

.9-.75（m，9H）

3-ethyl hexanoyl butyl NMR(DMSO-d ₆): 8.28(d,

J＝8，1H），8.14-8.04（m，

2H），4.34-4.10（m，2H），

4.10-3.95（m，2H），

3.75-3.62（m，2H），2.19

（t，J＝6，2H），2.04（d，

J＝6，2H），2.04-1.60

（m，3H），1.60-1.45（m，

2H），1.40-1.10（m，

13H），0.90-0.76（m，9H）

Embodiment 17

Repeat the method for embodiment 15 again, begin, just do not carry out hydrogenation, prepared following compounds with suitable reagent:

R ₁R ₄R ₅Nucleus magnetic resonance

3-ethyl hexanoyl butyl butyl NMR(DMSO-d ₆): 8.27(d, J=7,

1H），8.20（d，J＝7，1H），8.07

（t，J＝7，1H），4.37-4.13（m，

2H），4.02（t，J＝6，4H），

3.80-3.62（m，2H），2.20

（t，J＝6，2H），2.05（d，J＝6，

2H），2.02-1.64（m，3H），1.60-

1.47（m，4H），1.40-1.13（m，

13H），0.95-0.77（m，12H）

3-ethyl oenanthyl butyl butyl NMR(DMSO-d ₆): 8.27(d, J=7,

1H），8.20（d，J＝7，1H），8.06

（t，J＝6，1H），4.36-4.13（m，

2H），4.02（t，J＝6，4H），

3.80-3.60（m，2H），2.20

（t，J＝6，2H），2.04（d，J＝6，

2H），2.00-1.60（m，3H），1.60-

1.49（m，4H），1.40-1.10（m，

15H），0.95-0.72（m，12H）

R ₁R ₄R ₅Nucleus magnetic resonance

3-methyl oenanthyl butyl butyl NMR(DMSO-d ₆): 8.26(d, J=7,

1H），8.19（d，J＝7，1H），8.07

（t，J＝6，1H），4.32-4.11（m，

2H），4.02（t，J＝5，4H），

3.79-3.59（m，2H），2.20

（t，J＝6，2H），2.14-1.68（m，

5H），1.61-1.46（m，4H），1.40-

1.06（m，13H），0.95-0.81（m，

12H）

3-S-benzyl benzyl NMR(DMSO-d ₆): 8.33(d, J=7,

Methyl oenanthyl 1H), 8.24(d, J=7,1H), 8.08

（t，J＝5，1H），7.33（s，10H），

5.08（s，4H），4.40-4.22（m，

2H），3.80-3.60（m，2H），2.21

（t，J＝5，2H），2.14-1.64（m，

5H），1.26（d，J＝7，3H），

1.22-0.98（m，6H），0.88-0.73

（m，6H）

Embodiment 18

N-(3-(S)-methyl oenanthyl-D-gamma-glutamyl-glycyl-D-L-Ala crystallization

N-(3-(S)-methyl oenanthyl-D-gamma-glutamyl (α-benzyl ester)-glycyl-D-L-Ala benzyl ester (30.8 gram) is mixed into slurry in 2 liters of autoclaves with in 300 milliliters of dehydrated alcohols.Add 5%Pd/C1.54 gram (50% water is moistening), mixture carries out hydrogenation 1 hour under 4 normal atmosphere, inhales hydrogen therebetween promptly fully.Filtering recovering catalyst filters with paper for the first time, filters on 0.45 micrometer nylon micro pore sheet then; shift and washing catalyst with 100～150 milliliters of ethanol, the filtrate of this merging and washings are concentrated obtain moist white solid, it is dissolved in the mixture of 1: 10 dehydrated alcohol of 150 milliliters of heat and acetonitrile; filtered while hot is clarified, and boiling is concentrated to 35 milliliters, is cooled to room temperature gradually and makes it granulation and filtration; it is intensive to obtain crystalloid, and non-electrostatic title compound 20.1 grams (94%), its infrared spectra of making of paraffin oil have following main; differentiate specific absorption very clearly: 3340; 3300,2900,2836; 1725; 1650,1628,1580; 1532; 1455,1410,1370; 1280; 1240,1216, and 1175cm ^-1

This crystalline product (9.4 gram) is dissolved in 1000 milliliters the acetone and refluxed 1 hour, can be further purified.Solution is chilled to room temperature and adds a little above-mentioned crystallization as seed crystal, stirred 6 hours, by filter with a spot of acetone of trying one's best, washing, in 35 ℃ of dried recovered title product of vacuum, 7.25 grams, it has identical infrared spectral characteristic.

Embodiment 19

N-(3-(R)-methyl-4-heptene acyl)-D-gamma-glutamyl (α benzyl ester)-glycyl-D-L-Ala benzyl ester

Press the step of embodiment 10D, from D-gamma-glutamyl (the α benzyl ester)-glycyl-D-L-Ala benzyl ester hydrochloride of 2.77 grams (5 mmole) with from the 3-(R of 747 milligrams (5 mmoles))-acyl chlorides of methyl-4-heptenoic acid preparation makes title compound.

Embodiment 20

N-(3-(S)-methyl-4-oenanthyl)-D-gamma-glutamyl-glycyl-D-L-Ala

The mixture of 500 milligrams and the 26 milligram 5% palladium charcoal of product (50% water-wet) that obtains with embodiment 19 is jolting 2.5 hours in the 4 atmospheric nitrogen atmosphere in 125 milliliters ethanol, at initial pressure.The elimination catalyzer, removal of solvent under reduced pressure, product passes through the method purifying of embodiment 18, and all just the same in every respect with this routine product.

Preparation method A

The cyclohexyl Acetyl Chloride 98Min.

A1. cyclohexyl vinyl acetic monomer

Enough hexanes are added in 60% sodium hydride of 4.9 grams in oil; with dissolving oil; under nitrogen, in the sodium hydride that does not have oil, add 100 milliliters of exsiccant tetrahydrofuran (THF)s, then add 80 milliliters of dry tetrahydrofuran solution that contain 22.2 milliliters of phosphono acetic acid triethyls again.After 1 hour, add 40 milliliters of tetrahydrofuran solutions that are dissolved with 10.5 milliliters of pimelinketone in stirring at room, this reaction mixture in stirred overnight at room temperature, is poured into this reaction mixture in the water again, use extracted with diethyl ether.Organic phase 1N sodium hydroxide solution, water and salt water washing.Separate organic phase, use dried over mgso, and concentrating under reduced pressure.

Resistates is dissolved in 250 ml methanol, handle with 10% palladium hydroxide that 1.5 grams are stated from the carbon, with mixture in 3.52 kilograms per centimeter ²Hydrogen pressure under jolting 4 hours.The elimination catalyzer, filtrate decompression concentrates.Resistates distills under 45～50 ℃/0.4 holder, gets the purpose intermediate of 15.4 grams (yield 90%).

A2. cyclohexyl Acetyl Chloride 98Min.

The potassium hydroxide of 15.2 grams are added in 100 milliliters of methyl alcohol that contain 15.4 gram cyclohexyl vinyl acetic monomers, this solution was refluxed 3 hours.Pressure reducing and steaming methyl alcohol, resistates is with water treatment.The aqueous solution is used 10% hcl acidifying then with extracted with diethyl ether.This acidifying aqueous solution extracts with fresh ether, then, separates organic phase, water and salt water washing, and dry back is removed and is desolvated, and obtains liquid residue.

Resistates is dissolved in 60 milliliters the methylene dichloride and and handles with 18 milliliters oxalyl chloride.After the stirring at room 4 hours, with the reaction mixture concentrating under reduced pressure, and distillation residue, product distillates in 45～50 ℃/0.4 holder, meter 12.5 grams (yield 86%).

Preparation method B

By the general operation of preparation A with triethyl phosphine acyl group acetal and suitable aldehyde and ketone as starting raw material, the chloride of acid below having prepared:

R ₁COCl

R ₁Boiling point ℃/torr

（CH ₃CH ₂CH ₂） ₂CHCH ₂- 50-55/0.4

（CH ₃CH ₂） ₂CHCH ₂- 22-25/0.5

23-30/0.5

22-25/0.5

（CH ₃） ₂CH（CH ₂） ₃- 24-31/0.7

R ₁Boiling point ℃/torr

34-37/0.5

45-47/0.6

25-30/0.5

（CH ₃CH ₂） ₂CH（CH ₂） ₂- 32-36/0.4

30-38/.06

63-65/.95

89-92/5

46-50/0.5

R ₁Boiling point ℃/torr

30-34/0.5

31-35/0.7

Preparation method C

6-methyl oenanthyl chloro

C1.3-hydroxy-4-methyl-1-amylene

Be added drop-wise in the solution that the 1.0 molar ethylene base magnesium bromides that are as cold as 5 ℃ form in 90 milliliters of tetrahydrofuran (THF)s being dissolved in 6.3 milliliters of isobutyric aldehydes in 30 milliliters of tetrahydrofuran (THF)s, make this mixture be warming to room temperature then.After 2 hours reaction mixture is added in the saturated ammonium chloride solution, uses extracted with diethyl ether.Merge ether extraction liquid with saturated ammonium chloride solution, saturated sodium bicarbonate solution and salt water washing, use dried over mgso.Removal of solvent under reduced pressure gets 6.0 gram purpose products.

C2.6-methyl-4-oil of cognac

The mixtures of 18.2 gram 3-hydroxy-4-methyl-1-amylenes, 200 milliliters of triethly orthoacetates and 500 milliliters of right-toluenesulphonic acidss with 400 milliliters of O for toluene, and with 4A molecular sieve reflux 24 hours.Removal of solvent under reduced pressure, distillation residue.Fractionation under 45～64 ℃/0.5 holder gets 7.5 gram purpose products.

C3.6-methyl enanthic acid ethyl ester

In 7.5 gram 6-solution that methyl-the 4-oil of cognac forms, add 700 milligrams of palladium hydroxides that are stated from 10% on the carbon in 75 ml methanol, with this mixture in 3.52 kilograms per centimeter ²Hydrogen pressure under jolting 1.5 hours, elimination catalyzer, removal of solvent under reduced pressure, 5.7 gram purpose products.

C4.6-methyl oenanthyl chloro

By the working method of preparation A2, with the 6-methyl enanthic acid ethyl ester of 5.7 grams, obtain the purpose product of 2.0 grams, its boiling point is 30～34 ℃/0.5 holder.

Preparation method D

2-methyl oenanthyl chloro

The D1.2-methyl enanthic acid

With 5.4 milliliters just-enanthic acid be added to cold (0 ℃) by 11.8 milliliters of dry diisopropylamines and 55 milliliters of 1.6M concentration just-solution that butyllithium forms in 100 milliliters of dry tetrahydrofuran in, this mixture was stirring at room 1 hour.Gained solution is as cold as 0 ℃, adds 7.2 milliliters methane again.Reaction mixture stirred 1.5 hours under room temperature, nitrogen, poured into then in 10% hydrochloric acid, with ether (3 * 100 milliliters) extraction.Combining extraction liquid is used dried over mgso with 10% hydrochloric acid, water, 20% sodium bisulfite and salt brine solution washing, removal of solvent under reduced pressure, and 5.61 gram resistatess of gained are dissolved in the methyl alcohol that contains 5.1 gram potassium hydroxide.After stirring is spent the night, remove methyl alcohol, resistates is dissolved in 150 ml waters.With ether (2 * 100 milliliters) washing water layer, and use 10% hcl acidifying.Product (water layer) is used extracted with diethyl ether, and extraction liquid is used dried over mgso with 20% sodium sulfite solution and salt water washing.Remove ether, get the yellow liquid products of 5.0 grams.

D2.2-methyl oenanthyl chloro

With 5 gram 2-methyl enanthic acids and 7.6 milliliters of oxalyl chloride reactions, with the method for preparing A2, obtain the purpose product of 3.3 grams, its boiling point is 32～34 ℃/0.6 holder.

Preparation method E

3-(S)-the methyl oenanthyl chloro

E1.3-(R)-methylglutaric acid monomethyl ester

In 5 liters of four neck flasks of being furnished with agitator and pH electrode, add 2.5 liters of 0.01 mole of pH values and be 7 potassium hydrogen phosphate damping fluid, then add 150 milligrams of Pig Liver Esterases and 150 gram 3-methylglutaric acid dimethyl esters.The pH value of mixture maintains about 6.85 by 10% solution of potassium carbonate that regularly adds.2.5 after hour with 10% hydrochloric acid acidizing reaction liquid to pH be 2.0, use the extracted with diethyl ether product.Merge the extract dried over mgso, concentrating under reduced pressure gets 114 gram purpose products, (α) _D=-1.48(CH ₃The OHC=0.086 grams per milliliter).

E2.3-(R)-methyl-5-hydroxypentanoic acid methyl ester

114 gram 3-(R in being as cold as 0 ℃ 715 milliliters dry tetrahydrofuran)-methylglutaric acid monomethyl ester in, the concentration that slowly adds 391 milliliters is the borane dimethyl sulphide of 2M in tetrahydrofuran (THF).After adding, with reaction mixture in stirred overnight at room temperature.The cooling reactant slowly adds 50 ml waters.With ether (3 * 100 milliliters) extractive reaction mixture, combining extraction liquid water, saturated sodium bicarbonate solution and salt brine solution wash in succession, use dried over mgso, remove and desolvate, and get 37 gram purpose products.

E3.3-(R)-and methyl-5-(tertiary butyl dimethyl-siloxy-) methyl valerate

At 500 milliliters of dimethyl formamides and 37 gram (0.253 mole) 3-(R)-adding 37 gram (0.249 mole) tert-butyl dimethylsilane chlorine in the solution that the imidazoles of methyl-5-hydroxypentanoic acid methyl esters and 37 grams (0.543 mole) forms, reaction mixture was stirring at room 2 hours.Reaction mixture is poured in the water also with ether (4 * 100 milliliters) extraction.The extraction liquid that merges washs in succession with 10% hydrochloric acid, saturated sodium bicarbonate solution, water and salt brine solution, uses dried over mgso.Remove and desolvate, get 121.88 gram raw product, with its distillation, obtain 107.12 gram straight products, boiling point 80～81 ℃/0.4 holder.

E4.3-(S)-methyl-5-(tert-butyl dimethylsilane oxygen base)-the 1-amylalcohol

53.5 gram (0.206 mole) 3-(R in 125 milliliters of ether)-methyl-5-(tert-butyl dimethylsilane oxygen base) methyl valerate is added under nitrogen chlorine protection in 250 milliliters of ether that contain 8.5 gram (0.224 mole) lithium aluminium hydride.Reaction mixture stirred 1 hour in the time of 0 ℃, splashed into 8.4 gram water, 8.4 milliliter of 15% sodium hydroxide solution and 25.2 ml waters then.The elimination solid separates organic phase and water, 2.5% hydrochloric acid and salt brine solution washing, the organic phase dried over mgso, and concentrating under reduced pressure gets 46 gram products.

E5.3-(R)-methyl-5-(t-butyldimethylsilyloxy base)-the 1-valeral

74.81 gram dimethyl sulfoxide (DMSO) in 100 milliliters of dry methylene chloride are added drop-wise under nitrogen atmosphere in the 300 milliliters of dry methylene chloride that contain 56.3 gram oxalyl chlorides that are chilled to-60 ℃.After 15 minutes, drip the 3-(S of 250 milliliters of 92.0 grams in same solvent)-methyl-5-(tert-butyl dimethylsilane oxygen base)-the 1-amylalcohol.Add triethylamine after 30 minutes to-60 ℃ of temperature, then remove cryostat.Reaction mixture at room temperature stirred 1.5 hours, poured into then in the water, with dichloromethane extraction, extraction liquid was washed in succession with 2.5% hydrochloric acid, saturated sodium bicarbonate solution, water and salt brine solution, used dried over mgso then.Remove and desolvate, resistates is dissolved in ether washing and dry as in the previous once more.Remove ether, get 90.9 gram purpose products.

E6.5-(S)-and methyl-7-(tert-butyl dimethylsilane oxygen base)-2

-heptene

800 milliliters of dry tetrahydrofuran and 80 gram (0.2155 mole) triphenyl-ethyl bromo-phosphoniums are mixed into pulpous state, toward wherein add 165.7 ml concns be 1.3M in solvent equally just-butyllithium (0.2155 mole) solution.After 2 hours, drip 45 gram (0.196 mole) 3-(R in 200 milliliters of dry tetrahydrofuran in the reaction mixture)-methyl-uncle 5-(-dimethylsilane oxygen base)-the 1-valeral.Reaction mixture is poured in the water then stirring at room 2 hours, uses extracted with diethyl ether.Extraction liquid water that merges and salt brine solution washing, and use dried over mgso.Removal of solvent under reduced pressure, a yellow oil, through distill 37.4 gram products, boiling point 74～79 ℃/0.2～0.1 holder.

E7.3-(S)-methyl isophthalic acid-enanthol

Containing 74.8 gram 5-(S)-methyl-7-(tert-butyl dimethylsilane oxygen base)-500 ml methanol of 2-heptene in, add the palladium hydroxide that 7.5 grams are stated from 10% on the carbon.This mixture is in 3.52 kilograms per centimeter ²The down middle jolting of hydrogen pressure 1.5 hours.The elimination catalyzer, the pressure reducing and steaming solvent gets 30 gram products.

E8.3-(S)-methyl enanthic acid

In 45 minutes, at 15～20 ℃, toward containing 10 gram 3-(S)-175 milliliters of acetone of methyl isophthalic acid-enanthol in, drips 90 milliliters of Jones reagents, after 15 minutes, add 15 milliliters of Virahols again, and continue stirring 30 minutes.Reaction mixture is poured in the water into the product extracted with diethyl ether.Combining extraction liquid, water, sodium sulfite solution and salt brine solution wash in succession, and use dried over mgso.Remove and desolvate, get 10 gram product liquids.Boiling point 84～88 ℃/0.4 holder (α) _D=-4.46(CH ₂OH C=0.105 grams per milliliter)

E9.3-(S)-the methyl oenanthyl chloro

By the operation of preparation A2 by 5.0 gram 3-(S)-methyl enanthic acid and 7.5 milliliters of oxalyl chlorides can make the purpose chloride of acid of 2.9 grams, boiling point 29～32 ℃/0.25 holder.

Errata

After the preceding revisal of the capable revisal of file name page or leaf

Claims 1 fall 5 mutually separately

The subsalt subsalt product of 24 products

3 fall 5

27 to remove adjacent benzyl by catalytic hydrogenolysis be by catalysis hydrogen

Going the 0-benzyl to separate removes

32 is independently independently

The adjacent benzyl of the 2 0-benzyls that fall

Specification sheets 9 is 12 method-N-first

Fall 9 phenylisoxazlene phenyl Yi Er oxazole alkene

33 fall 7 pairs be dissolved in 150 milliliters of dioxs

{。##.##1},

Fall 7 to be dissolved in and to add 150 milliliters in 150 milliliter two and use hydrochloric acid

Fall 6 alkane ... saturated, saturated De diox,

35 propyl carbinols that fall 5 (0.516 moles) and (0.516 mole)

And

36 fall 5 ethanol methyl alcohol

53 fall 3 oenanthyl heptene acyls

57????11

Fall 2 enanthic acid heptenoic acids

Fall 7 orthoformic acid ortho-acetic acids

Errata

After the preceding revisal of the capable revisal of file name page or leaf

Specification sheets 53,5

6，9，

10，11

13，16????I????1

Fall 2

9 fall 4

7????15????Pergamn????Pergaman

9????13????method????metho

10 6 CH ₃C ₃H

55????14

56 1 boiling points ℃/boiling point ℃/holder

57????1

56 fall 3

59 5 150 milliliters 150 milligrams

Claims (6)

1, a kind of method for preparing following formula: compound,

In the formula: R ₁For the alkyl that contains 2 to 10 carbon atoms, contain the cycloalkyl of 4 to 7 carbon atoms or contain the methyl cycloalkyl of 6 to 8 carbon atoms; R ₂For hydrogen or contain the alkyl of 1 to 3 carbon atom; R ₃For hydroxyl or have an amino-acid residue of following formula,

In the formula: X is hydrogen, contains the alkyl or the methylol of 1 to 2 carbon atom that n is an integer of 0 to 4; R ₄For hydrogen, contain the alkyl or the benzyl of 1 to 6 carbon atom; And R ₅For hydrogen, contain the alkyl of 1 to 6 carbon atom, the methyl cycloalkyl or the benzyl of 6 to 8 carbon atoms, it is characterized in that, with having the compound of following formula,

R in the formula ₁And R ₂Definition as above and has following formula: compound dehydration coupling,

Wherein, the functional group that does not participate in reacting can be protected, and X, n and R ₅As defined above, remove blocking group subsequently, and if desired, preparation can be used for the subsalt of the product of medicine.

2, according to the process of claim 1 wherein that said coupling is to carry out in the presence of N-maloyl imines or I-hydroxybenzotriazole and carbodiimide.

3, according to the method for claim 2, wherein, the 0-benzyl in the protected group is removed by catalytic hydrogenolysis.

4, a kind of preparation has the method for the compound of following formula,

In the formula, R ₁For the alkyl that contains 2 to 10 carbon atoms, contain the cycloalkyl of 4 to 7 carbon atoms or contain the methyl cycloalkyl of 6 to 8 carbon atoms; R ₂For hydrogen or contain the alkyl of 1 to 3 carbon atom; R ₃For hydroxyl or have a kind of amino-acid residue of following formula,

In the formula, X is hydrogen, contains the alkyl or the methylol of 1 to 2 carbon atom that n is an integer of 0 to 4; And

R ₄For hydrogen, contain the alkyl or the benzyl of 1 to 6 carbon atom; And R ₅Be hydrogen, contain 1 to 6 carbon atom alkyl, contain the methyl cycloalkyl or the benzyl of 6 to 8 carbon atoms, it is characterized by: with compound with following formula

Compound with following formula is carried out acidylate,

In the formula, R ₂, R ₃, R ₅Define as above, at R ₁The functional group that does not participate among the CO-Hal reacting can be protected, R ₁As defined above, Hal represents a kind of halogen, removes protecting group subsequently, and if desired, preparation can be used for the subsalt of pharmaceutical prod.

5, according to the method for claim 4, wherein, acidylate is carried out having in the presence of the tertiary amine by acyl chlorides.

6, according to the method for claim 5, the 0-benzyl in the wherein protected group is removed by catalytic hydrogenolysis.