CN1096292A - The pyrrolidine derivatives as HIV protease inhibitors that replaces - Google Patents

Abstract

The present invention is about the compound of formula I:

Wherein X is a terminal group, for example, and aryloxy carbonyl or alkyloyl; B does not exist or is amino-acid residue, for example, and Xie Ansuan or aspartic acid; R ¹Be alkyl; And Y is a cyclic substituents, for example, and benzyl, benzyloxy, thiophenyl or 2-pyridine sulfenyl.The human cell's that this compounds inhibition HIV (human immunodeficiency virus) (HIV) protease activities and interference factor HIV cause cytopathy.These character make this compounds become the useful medicine that infects antagonism with HIV.

Description

The pyrrolidine derivatives as HIV protease inhibitors that replaces

The method that the invention relates to the preparation of the method that shows active compound to resisting special retrovirus, produce these compounds, medicine and utilize these compounds to go and struggle by the caused infection of retrovirus.

Nineteen eighty-three, a kind of retrovirus that is called as HIV (human immunodeficiency virus) the 1st type (HIV-1) is confirmed as the pathogeny body of Acquired immunity deficiency symptoms (AIDS), see R.C.Gallo and L.Montagnier, Scientific American(Scientific Beauty compatriots), 259(4), 40(1988).This virus has become a kind of eqpidemic disease with surprising ratio.Recently, closely-related virus, HIV (human immunodeficiency virus) the 2nd type (HIV-2) have been accredited as the 2nd kind of pathogeny body of AIDS.

As the identification of the HIV (human immunodeficiency virus) (HIV) of pathogeny body and the development of this viral method of a large amount of breedings, caused the discovery that can suppress the compound that HIV duplicates external.The most important classification of Shi Bie inhibitor compound is that a group belongs to the dideoxyribonucleoside compounds after this manner, wherein 3 '-azido--3 '-deoxythymidine (be also referred to as benefit 5 pyridines (Zidovndine) or AZT) and recently 2 ', 3 '-dideoxyinosine (is also referred to as Di Duonuoxin (didanosine) or DDI) is used for treating the patient that some has the HIV infection symptoms.Have found that this compounds is by suppressing the life cycle of reversed transcriptive enzyme interference HIV.This kind of enzyme is converted into double-stranded thymus nucleic acid (DNA) to viral RNA, and itself is a kind of indispensable enzyme that HIV duplicates.Except suppressing reversed transcriptive enzyme, other stages of HIV life cycle have also done bright, become the target of the anti-AIDSO medicine of development.A target that more and more comes on the scene is a kind of HIV-codase that is called as hiv protease.This kind of enzyme, resemble reversed transcriptive enzyme, by the pol genes encoding, and growth is essential for HIV, it is responsible for making gag(p55) or gag-pol(p180) produce some division in the protein and discharge structural protein, for example p17 and p24, and enzyme, comprise itself, these enzymes can find in the virus particle of sophisticated infection.Therefore, the inhibitor of hiv protease can be blocked the life cycle of HIV.

Can from the increase of the drug report of finding blocking-up this kind of enzyme reflect for growing concern that hiv protease gave recent years.For example, can see the up-to-date review of writing by D.W.Norbeck and D.J.Kempf, step on Chemistry at Annual Report In Medicinal about the inhibitor aspect of proteolytic enzyme, 26,141(1991) on.(deliver as in nearest review, mentioning by people such as D.H.Rich, step at J.Med.Chem.33,1285(1990) and people such as N.A.Roberts deliver, step at Science, 248,358(1990) on), in peptide with p17/p24 substrate breaking point sequence, by the replacement of ethylol amine transition state analogues (TSA), can obtain the series product of two kinds of effective hiv protease inhibitor.The biological study paper of people's such as Roberts guiding compound is delivered by people such as H.A.Overton, step at Virology, 179,508(1990), by stepping on that people such as J.A.Martin deliver at Biochem, Biophys.Res.Commun., 176,180(1991) go up and step on andChemotheraphy at Antiviral Chemistry by what people such as J.C.Craig delivered, 2,181(1991) on.

Other is disclosed to have the comprising of hiv protease inhibitor of ethylol amine TSA:

People such as B.K.Handa, No. the 346847th, European patent application, 1989,12,20 is open.

People such as G.B.Dreyer, No. the 352000th, European patent application, 1990,1,24. is open.

People such as D.J.Kempf, No. the 402646th, European patent application, 1990,12,19. is open.

With people such as K.E.B.Parkes, the Canadian patent application case, the 2nd, 030, No. 415,1991,6,12. is open.

J.A.Martin and S.Redshaw, European patent application, No. 432695,1991,6,19. is open.

The invention describes the pyrrolidin derivatives of replacement, these derivatives have in the structure that ethylamine TSA is combined in them.These derivatives all are effective inhibitor of hiv protease.Especially these compounds have the cytopathic ability that suppresses the human body cell that HIV causes and are demonstrated.These character, the attribute of relative selectivity effect and significantly nontoxicity in addition become with HIV this compounds and infect right useful medicine.

Compound of the present invention is as shown in Equation 1:

Wherein:

X is R ²OC(O), R ²C(O) or R ²NR ²C(O), R wherein ²For

(ⅰ) low-carbon alkyl,

(ⅱ) low-carbon naphthenic,

(ⅲ) phenyl or with the mono-substituted phenyl of halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy,

(ⅳ) phenyl (low-carbon (LC)) alkyl or

Phenyl (low-carbon (LC)) alkyl, wherein

Its aromatic base part replaces with halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy list,

(ⅴ) 1-naphthyl or 2-naphthyl,

(ⅵ) (Het) or (Het)-(low-carbon alkyl), wherein

Het represents 5 or 6 yuan, contains 1 or 2 unit price heterocyclic radical that is selected from the heterocyclic atom of nitrogen, oxygen and sulphur, or

(ⅶ) 2-quinolyl or 3-quinolyl, and R ²Be hydrogen or low-carbon alkyl;

Or X is R ^2AOCH ₂C(O), R wherein ^2AFor phenyl or with low-carbon alkyl or halogen

Mono-substituted, disubstituted or trisubstd phenyl;

B is not for existing or divalent radical-NHCHR ⁴C(O)-, R wherein ⁴Be low-carbon alkyl; Low-carbon naphthenic; (low-carbon naphthenic)-(low-carbon alkyl); Phenyl methyl; Or with hydroxyl, carboxyl, lower alkoxycarbonyl, aminocarboxyl, (low-carbon alkyl) aminocarboxyl or two (low-carbon alkyl) aminocarboxyl;

R ¹Be low-carbon alkyl or low-carbon naphthenic;

Y is a low-carbon alkyl; Low-carbon naphthenic; Phenyl or with halogen, the mono-substituted phenyl of hydroxyl, low-carbon alkyl or low-carbon alkoxy; Phenyl methyl or with the mono-substituted phenyl methyl of halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy; Or

Y is W(CH ₂)

Z, wherein W is oxo, sulfo-, sulfinyl or sulfonyl, Z is a low-carbon alkyl; Phenyl or with the mono-substituted phenyl of halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy; Or (Het), wherein (Het's) is described as defined above; And n is 0 or 1;

Or formula (1) compound acceptable acid addition salt in treatment.

About term in the formula 1 " B is not for existing ", expression symbol B has become " X " has been connected to covalence key on this secondary amino group, otherwise this amino will be connected to " B ".

Represent one group of preferable The compounds of this invention with formula 1, wherein X is R ²OC(O) or R ²C(O), R wherein ²Be low-carbon alkyl; Phenyl (low-carbon (LC))-alkyl; Phenyl

(low-carbon (LC)) alkyl, wherein the position 4 of phenyl moiety is replaced by chlorine, fluorine, hydroxyl, methyl or methoxy; The 1-naphthyl; The 2-naphthyl; The 2-furyl; The 2-thienyl; The 2-pyridyl; The 4-pyridyl; 2-pyridyl, methyl; 4-thiazolyl methyl or 2-quinolyl; Or

X is R ^2AOCH ₂C(O), R wherein ^2ABe phenyl

Or do single, double or trisubstd phenyl in the position that is selected from position 2,4 and 6 with low-carbon alkyl or halogen;

B is not for existing or divalent radical-NHCHR ⁴C(O)-, R wherein ⁴Be low-carbon alkyl, or with the mono-substituted low-carbon alkyl of hydroxyl, low-carbon alkoxy carbonyl, aminocarboxyl, (low-carbon alkyl) aminocarboxyl or two (low-carbon alkyl) aminocarboxyl;

R ¹Be the 1-methylethyl, 1,1-dimethyl ethyl, 2-methyl-propyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

Y is low-carbon naphthenic, phenyl, 4-chloro-phenyl-, 4-bromophenyl, 4-fluorophenyl, 4-tolyl, 4-methoxyphenyl, phenmethyl, (4-fluorophenyl) methyl or (4-tolyl) methyl; Or

Y is W(CH ₂)

Z, wherein W and n's is described as defined above, and Z is low-carbon alkyl, phenyl, 2-furyl, 2-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 4-thiazolyl, 2-pyrimidyl, 4,6-dimethyl-2-pyrimidyl or 2,6-dimethyl-4-pyrimidyl;

Maybe should group preferred compounds acceptable acid addition salt in treatment.

Represent one group of better compound with formula 1, wherein X is uncle-butoxy carbonyl, carbobenzoxy-(Cbz), (4-chloro-phenyl) methoxycarbonyl, (4-hydroxyphenyl) methoxycarbonyl, (4-methoxyphenyl) methoxycarbonyl, ethanoyl, benzoyl, 1-naphthalene carbonyl, 2-naphthalene carbonyl, 2-pyridine methoxycarbonyl, 2-quinoline carbonyl, benzene oxygen ethanoyl, (2-tolyloxy) ethanoyl, (2, the 4-dimethyl phenoxy) ethanoyl, (2, the 6-dimethyl phenoxy) ethanoyl, (2,4,6-trimethylammonium phenoxy group) ethanoyl, (4-chlorophenoxy)-ethanoyl or (4-fluoro-2,6-dimethyl phenoxy) ethanoyl;

B is not for existing or being divalent radical-NHCHR ⁴C(O)-, R wherein ⁴Be the 1-methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 2-methyl-propyl, methoxycarbonyl methyl, ethoxycarbonylmethyl group or amino carbonyl methyl;

R ¹Be 1,1-dimethyl ethyl or cyclopropyl;

Y is a cyclohexyl, phenyl, 4-chloro-phenyl-, the 4-fluorophenyl, 4-methoxyphenyl, benzyl, (4-methoxyphenyl) methyl, 2-methyl propoxy-, phenoxy group, the 2-pyridyloxy, 3-pyridyloxy, 4-pyridyloxy, the 2-2-pyrimidinyl oxy, 4,6-dimethyl-2-2-pyrimidinyl oxy, 2,6-dimethyl-4-2-pyrimidinyl oxy, benzyloxy, 2-pyridine methoxyl group, 4-thiazolyl methoxyl group, 2-pyrimidine methoxyl group, thiophenyl, benzene sulfinyl, benzene sulfonyl, 2-pyridyl sulfenyl, 3-pyridyl sulfenyl, 4-pyridyl sulfenyl, the 2-pyrimidine-based sulfur-base, 4,6-dimethyl-2-pyrimidine-based sulfur-base, benzylthio-, benzyl sulfinyl, benzyl sulfonyl, (2-picolyl) sulfenyl, (3-picolyl) sulfenyl or (4-picolyl) sulfenyl; Maybe should the better compound of group acceptable acid addition salt in treatment.

Represent the compound of one group of the best with formula 1, wherein X is uncle's one butoxy carbonyl, carbobenzoxy-(Cbz), ethanoyl, 2-naphthyl carbonyl, 2-pyridine methoxycarbonyl, 2-quinoline carbonyl;

B is valyl, isoleucyl or asparagyl;

R ¹Be 1,1-dimethyl ethyl or cyclopropyl and

Y is a phenyl, benzyl, phenoxy group, 2-2-pyrimidinyl oxy, 2,6-dimethyl-4-2-pyrimidinyl oxy, benzyloxy, thiophenyl, benzene sulfonyl, 2-pyridine sulfenyl, 3-pyridine sulfenyl, 4-pyridine sulfenyl, 2-pyrimidine sulfenyl, 4,6-dimethyl-2-pyrimidine sulfenyl or (3-picolyl) sulfenyl; Maybe should group optimizing compound acceptable acid addition salt in treatment.

Represent the compound that another group is best with the formula I, wherein X is (2-methylphenoxy) ethanoyl, (2, the 4-dimethyl phenoxy) ethanoyl, (2, the 6-dimethyl phenoxy) ethanoyl or (2,4,6-trimethylammonium phenoxy group) ethanoyl; B is not for existing; R ¹Be 1, the 1-dimethyl ethyl; And the definition of Y is shown in above-mentioned last example; Or be this another group optimizing compound acceptable acid addition salt in treatment.

About the compound of formula 1, being preferably wherein, B is divalent radical-NHCHR ⁴C(O)-, this has R ⁴Asymmetric carbon atoms have (S) configuration.

Comprise within the scope of the present invention be a kind of pharmaceutical composition that human body HIV infects that is used for the treatment of, this pharmaceutical composition comprises the compound of formula 1, or in treatment its salt of acceptable, and pharmaceutically useful carrier.

Scope of the present invention also comprises a kind of method that human body HIV infects that is used for the treatment of, and this method comprises formula 1 compound of taking significant quantity, or in treatment acceptable its salt.

In this scope, also comprise a kind of pathogenetic method of human body cell opposing HIV that is used to protect, this method comprise with a kind of formula 1 compound of anti-HIV significant quantity or in treatment acceptable its salt, handle above-mentioned cell.

The method of description preparation formula 1 compound.

Usually, the abbreviation of amino acid and protecting group is indicated in used here confession, is based on the suggestion of IUPAC-IUB biochemical nomenclature commission, sees European Journal of Biochemistry 138,9, (1984).For example, Val, Ile, Asn, with Len represent respectively L-Xie Ansuan, L-Isoleucine, altheine, with the residue of the bright amino acid of L-.

Term used herein " low-carbon alkyl ", separately or and the atomic group combination, expression contains the straight chained alkyl group of 1 to 6 carbon atom and contains the branched alkyl group of 3 to 4 carbon atoms and comprise methyl, ethyl, propyl group, butyl, hexyl, the 1-methylethyl, the 1-methyl-propyl, 2-methyl-propyl and 1, the 1-dimethyl ethyl.

Term used herein " low-carbon naphthenic ", separately or and the atomic group combination, expression contains the saturated cyclic hydrocarbon radical from 3 to 6 carbon atoms, and comprises cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

Term used herein " low-carbon alkoxy " expression contains the straight chain alkoxyl group of 1 to 6 carbon atom, contain the branched alkoxy of 3 to 4 carbon atoms and comprise methoxyl group, oxyethyl group, propoxy-, hexyloxy, the 1-methyl ethoxy, butoxy and 1,1-dimethyl oxyethyl group.Back one group is commonly referred to uncle's one butoxy.

Term used herein " halogen " expression is selected from the halogen radical of bromine, chlorine, fluorine or iodine.

Term used herein " Het " expression is removed a hydrogen atom and deutero-unit price base from the saturated or unsaturated heterocycle of 5-or 6-unit, these 5 or 6 yuan of saturated or unsaturated heterocycles contain 1 to 2 heteroatoms that is selected from nitrogen, oxygen and sulphur.This heterocycle can randomly have 1 or 2 substituting group; For example, low-carbon alkyl, low-carbon alkoxy, halogen, amino or low-carbon alkyl amino.The heterocycle that is fit to comprises tetramethyleneimine, tetrahydrofuran (THF), thiazolidine, pyrroles, 1H-imidazoles, 1-methyl isophthalic acid H-imidazoles isoxazole, thiazole, 2-methylthiazol with the heterocyclic example that can choose replacement wantonly, thiazolamine, piperidines, 1, the 4-diox, 4-morpholine, pyridine, the 2-picoline, pyrimidine, 4-methylpyrimidine and 2,4-dimethyl pyrimidine.

About amino acid whose term " residue ", represent self-corresponding a-amino acid by removing a hydroxyl on the decarboxylate and an alpha-amino hydrogen atom deutero-base.

Term used herein " pharmaceutically useful carrier " expression is a kind of nontoxicity, is generally the inert carrier for active ingredient, and it does not have an adverse influence to active ingredient.

The predetermined amount of term used herein " significant quantity " expression compound of the present invention enough resists intravital HIV effectively.

Method

Usually, the compound of formula 1 uses the known reaction conditions that is suitable for reactant to prepare by known method.Preparation method's description can be found in standard textbook, standard textbook such as " Annual Report In Organic Systheesis-1990(" annual report of organic synthesis-1990 ") people such as K.Turnbull, Eds; university press company, San Diego, California; the U.S., 1990(and previous annual report), " Vogel ' s Textbook of Practical Orgainic Chemistry " (" fertile outstanding practical organic chemistry textbook "); people such as B.S.Furniss, Eds, gantry Group Co.,Ltd; Ai Saisi economizes; Britain, 1986, with " The Peptides:Analysis, Synthesis, Biology " (peptide: analyze, synthetic, biology "), people such as E.Grass, Eds, university press, New York, the New York, the U.S., 1979-1987, the 1st to 9.

In further detail, the method for preparation formula 1 compound comprises:

(a) with the epoxide of formula 2, wherein the definition of X is as described herein

Tetramethyleneimine carboxylic acid amides reaction with formula 3

R wherein ¹As described herein with the definition of Y, can obtain the corresponding compound of formula 1, X wherein, R ¹As described herein with the definition of Y, and B is not for existing; Or

(b) with the compound of formula 4

R in the formula ²The activity derivatives reaction of and carboxylic acid X-OH as described herein with the definition of Y, wherein X is R ²C(O) or R ^2A

OCH ²C(O), its definition is as described herein, can obtain the corresponding compound of formula 1, and wherein X is R ²C(O) or R ^2AOCH ₂C(O), its definition as

Described herein, R ¹As described herein with the definition of Y, and B is not for existing;

(c) under the situation that coupling agent exists, coupling formula 4 compounds, wherein R ¹As described herein and the formula X-NHCHR with the definition of Y ⁴The a-amino acid of COOH, wherein X and R ⁴Definition as described herein, can obtain the corresponding compound of formula 1, wherein X, R ¹As described herein with the definition of Y, and B is divalent radical-NHCHR ⁴C(O)-, R wherein ⁴Definition as described herein, or

(d) with the compound of formula 5

R wherein ¹, R ⁴As described herein with the definition of Y, with the activity derivatives reaction of carboxylic acid X-OH, wherein X is R ²C(O) or R ^2AOCH ₂C(O), its definition is as described herein, can obtain the corresponding compound of formula 1, and wherein X is R ²C(O) or R ^2AOCH ₂C(O), its definition is as described herein, R ²As described herein with the definition of Y, and B is divalent radical-NHCHR ⁴C(O)-, R wherein ⁴Definition as described herein; And

(e) if needs are arranged, can with as by front (a) and (b), (c) or (d) compound of the formula 1 that obtained of part be transformed into acceptable acid addition salt in treatment.

Be noted that: the kind of formula 1 compound, wherein X is normally used N-protected base, Boc for example, Z, Fmoc or to a methoxy carbobenzoxy-(Cbz), usefulness method (a) and be very easy to (c) time and obtain expediently.This kind easy to be acquired becomes them and can be used as the intermediate of preferred routes, respectively via method (b) with (d), with the respective compound of production 1, wherein X is not normally used N-protected base.Therefore; as intermediate; the compound of this type of formula 1 will be gone protection (to that is to say; protecting group is removed), according to method (b) with (d) with the corresponding compounds of the terminal free amine of the N-of gained as formula 4 or formula 5, according to B for not existing or existing; with final preparation formula 1 compound; wherein X is not normally used N-protected base, for example, and 2-pyridine methoxycarbonyl or 2-quinoline carbonyl.

More clearly, according to the method (a) of front, B is the N-alkylated reaction preparation that non-existent formula 1 compound can add formula 3 tetramethyleneimine carboxylic acid amides by wushu 2 epoxide.In 20 °～110 ℃ temperature range, two kinds of reactants are contacted in the inert solvent as glycol, tetrahydrofuran (THF) or dimethyl formamide, this reaction is eligibly finished.Reaction times is depended on the character of temperature and reactant, usually in 2～24 hours scope.

According to method (b), the corresponding compounds of through type 4 and carboxylic acid X-OH(wherein X are respectively defined in this article R ²C(O) or R ^2AOCH ₂C(O) activity derivatives reaction, (wherein B does not exist, and X is defined R in this article can to obtain the compound of formula 1 ²C(O) or R ^2AOCH ₂C(O).Suitable reactive derivative is the acylating agent of the acyl group X-CO that can provide suitable, and comprises corresponding acid halide, preferentially selects muriate or bromide for use, active lipid, acid anhydride class or mixed anhydride class.With the condition of finishing reaction reaction is finished in accordance with known methods.These methods comprise by the ratio of selecting suitable reactant or if desired, by provide known protecting group to any other the active group with the active group competition of expectation temporarily, reach optionally method of desired response.Usually, temperature is 0 °～50 ℃, and the reaction times is finished this reaction in the inert solvent as tetrahydrofuran (THF), dimethyl formamide or methylene dichloride in 15 minutes to 24 hours scope.

According to method (C), under the situation that coupling agent exists, by the compound and the formula α-NHCHR of coupling formula 4 ⁴The a-amino acid of COOH, can obtain B is divalent radical-NHCHR ⁴C(O) compound of-formula 1 ,-NHCHR ⁴C(O)-in R ⁴As defined herein.Promote that with coupling agent the dehydration coupling of the free amino group of the free carboxyl group of a reactant and another reactant is that everybody knows; For example, see " peptide: analyze, synthesize biology ", the 1st to 8 volume, the source sees above.The suitable coupling agents example is 1,1 '-carbonyl-diimidazole or N, N '-dicyclohexyl-carbodiimide.Other example is at N, I-hydroxybenzotriazole or N-ethyl-N '-[(3-dimethyl-amino) propyl group] carbodiimide under N '-dicyclohexyl-carbodiimide exists.A kind of very actual and useful coupling agent be commercially available (the basic oxygen base of benzotriazole-1-) three-(dimethylamino) Phosphonium hexafluorophosphate, no matter be itself or in the presence of I-hydroxybenzotriazole.Another very actual and useful coupling agent is at the commercial 2-(1H-benzotriazole-1-yl that can obtain)-N, N, N ', N '-tetramethyl- A tetrafluoro borate.

Linked reaction is carried out in inert solvent, inert solvent such as methylene dichloride, acetonitrile or dimethyl formamide.Add excessive organic amine such as diisopropylethylamine or N-methylmorpholine, make the potential of hydrogen of reaction mixture maintain the pH value and be approximately 8.About-20 ° to 30 ℃, the reaction times was from 15 minutes to 8 hours usually for temperature of reaction.

Reference method (d), this method is carried out by the identical mode of method mentioned above (b), and unique exception is to use the compound of formula 5 to replace the compound of formula 4 as starting material.

Epoxide as the formula 2 of the starting material of method (a) all is known or can prepares with known method.More exactly, the epoxide of formula 2 is described by people such as B.K.Handa, No. the 346th, 847, European patent application, and 1989,12,20. is open, and perhaps the epoxide of formula 2 can be by the method manufacturing of describing in this patent application case.

Other starting material of these methods that is to say the tetramethyleneimine carboxylic acid amides of formula 3, and the compound with formula 4 and formula 5 is new compound, therefore, becomes one of purpose of the present invention.The appropriate methodology of preparation formula 4 and 5 compound is in above mentioning.By standard amination to known corresponding pyrrolidine carboxylic acid, tetramethyleneimine carboxylic acid amides that can preparation formula 3.Its also available F.Soucy, the method preparation of D.Wernic and P.Beaulieu.J.Chem.Soc.Perkins Trans.1，2885（1991）。The method of the tetramethyleneimine carboxylic acid amides of production formula 5 will illustrate in example hereinafter.

Formula 1 compound of the present invention can the treatment on acceptable acid addition salt form and obtain.The example of these salts is for having the organic acid salt, said organic acid such as acetate, lactic acid, succsinic acid, phenylformic acid, Whitfield's ointment, methylsulfonic acid or right-toluenesulphonic acids, also can be polymeric acid, as Weibull or carboxymethyl cellulose, with the salt that has mineral acid, said mineral acid such as haloid acid, for example hydrochloric acid, sulfuric acid, or phosphoric acid.If want, can be converted into another kind of acid addition salt to 1 concrete acid addition salt, for example nontoxicity, pharmaceutically useful salt, can be by with stepping at Helv.Chim.Acta, 43, the method that 1849(1960) goes up, described by people such as R.A.Boissonnas is with suitable ion exchange resin treatment.

Usually, the salt of the peptide of acceptable formula 1 is equivalent to these peptides itself fully biologically in treatment.

The compound of formula 1 or in treatment acceptable its hiv protease inhibition activity and the nosogenetic cytoprotection of anti-HIV of salt, can be by biochemical, microbiological and biological method explanation.

Be used for the compound of formula 1 or their a kind of useful especially method of the hiv protease inhibition activity of acceptable salt in treatment and be " Recombinant HIV proteolytic enzyme HPLC analytical method ".This method suppresses to have the ability of enzymic hydrolysis of hiv protease of the decapeptide (substrate) of aminoacid sequence based on test compound, this amino acid series contains the hiv protease breaking point of a known HIV polyprotein matter; See people such as H.G.Krausslich, Proc.Natl.Acad.Sci, USA, 86,807, (1989).Narrate in the detailed description of this analytical method and the result that example the obtained example hereinafter as formula 1 compound.

The compound of formula 1 or the ability that acceptable their salt protection cell resistance HIV infects in treatment, can to the human T cell be with the evaluation test compound in the pathogenic inhibiting microbiological method explanation of cell of HIV.The exemplary of these methods is described in S.Harada and N.Yamamoto, Jpn.J.Cancer Res.(Gann), 76,432(1985) and people such as S.Harada, and Science, 229,563(1985).Narrate in a kind of analytical method example hereinafter based on latter's method.

When compound of the present invention, or in treatment acceptable its salt, be used for infecting when struggling with the HIV of human body, can with a kind of vehicle that contains one or more pharmaceutically useful carriers through the oral cavity, epidermis or parenteral route introduce peptide, the ratio of various carriers depends on the solubleness and the chemical property of this compound, selected route of administration and the application of standard biologically.For oral administration, can with this compound or in treatment acceptable its salt be mixed with unit dosage form, for example capsule or tablet, in pharmaceutically useful carrier, each capsule or tablet approximately contain the activeconstituents of 5～150mg predetermined amount.For the epidermis administration, can with this compound with pharmaceutically useful, contain 0.01～2%, the preferably vehicle of 0.05～1% promoting agent preparation.This class prescription can be emulsifiable paste, lotion, the sublingual tablet preferably patch or the cheek patch of percutaneous dosing.For parenteral administration, with the form of the composition that has pharmaceutically useful vehicle or carrier, by the compound of vein, subcutaneous or intramuscular injection drawing-in system 1.For by drug administration by injection, it is preferable using the compound solution in the sterile liquid vehicle, and this vehicle also can contain other solution, and for example pharmaceutically useful salt or the glucose of buffer reagent or protective material and q.s are so that the solution isoosmotic pressure.

The vehicle or the carrier that are fit to that are used for above-mentioned prescription can find at the textbook of standard, for example, and at " Remington ' s Pharmaceutical Sciences ", 18th ed., Mack Publishing Company, Easton, Penn, 1990.(" the pharmacy science of Lei Mingdun; the 18th edition, Mike publishing company, Easton; Binzhou, 1990).

The dosage of compound changes with administering mode and selected concrete promoting agent.And, will become with the specific host under the treatment.Usually, treatment begins with low dose, is significantly less than the optimal dose of peptide, and thereafter, this dosage reaches best effect with little increasing amount increase until in this case.Usually, wish very much can not cause any concentration level disadvantageous or deleterious side effect to introduce this compound generally producing the result of antiviral effect.

For oral administration, every day per kilogram of body weight 5～150mg, preferable scope is per kilogram 5～50mg, introduce this compound or the treatment on acceptable salt.About the general administration, though above-mentioned changing conditions can take place, with the compound of dosage drawing-in system 1 of per kilogram of body weight 10 μ g～1000 μ g every day.

Infect for curing HIV, though above the various prescriptions of Jie Shiing all are effective and quite safe medicines, but take the possibility that can obtain favourable result when not getting rid of these prescriptions and other antiviral or medicament, other antiviral or medicament like this comprises soluble CD4, benefit 5 pyridines (Zidovudine), Di Duonuoxin (didanosine), Cha Xitabin (Zalcitabine), foscarnet sodium, Ribavirina (ribavarin), acycloguanosine (acyclovir), or antiviral Interferon, rabbit, (for example, alpha-interferon or interleukin-2(interleukin-2)).

Following example further specifies the present invention.Solution per-cent or ratio are represented the mutual relationship of volume to volume, unless otherwise mentioned.Temperature is represented with degree centigrade.On Bruker 200MHz spectrometer, write down proton magnetic resonance (PMR) (NMR) spectrum; With per hundred very much (PPM) write down this chemical shift (δ).The abbreviation of using in example comprises Boc: uncle-butoxy carbonyl; Bop:(benzotriazole-1-base oxygen base) three (dimethylamino) Phosphonium hexafluorophosphates; Bu: tert-butyl; BZL: benzyl; DIEA: diisopropyl ethyl amine; DMF: dimethyl formamide; HEPES:N-2-hydroxyethyl-piperazine-N '-2-ethylsulfonic acid; Et ₂O: ether; EtOAc: vinyl acetic monomer; EtOH: ethanol, HPLC: high efficiency liquid chromatography: MeOH: methyl alcohol; Ph: phenyl; THF: tetrahydrofuran (THF); Z: carbobenzoxy-(Cbz).

Example 1

4(S)-and benzyloxy-N-tert-butyl-uncle 1-(-butoxy carbonyl) preparation of tetramethyleneimine-2(S)-carboxylic acid amides.

(a) acid of N-protected; uncle 1-(-butoxy carbonyl)-the 4(S-hydroxyl pyrrolidine-2(S)-carboxylic acid; pass through 4(S)-hydroxyl pyrrolidine-2(S)-carboxylic acid is { suitable-4-hydroxyl-L-proline(Pro); described by S.G.Ramaswamy and E.Adams; step at J.Org.Chem.(organic chemistry periodical), 42,3440(1977) on and two-tert-butyl carbonate; in the presence of excessive NaOH, at THF/H ₂O(1: 1) in the solution under room temperature reaction prepared in 18 hours.

(b) in the acid of the N-protected that will obtain like this (400mg 1.73mmol) is dissolved in DMF(7ml).(99%, 87mg 3.63mmol) joins in this solution with sodium hydride.With this mixture (20～22 ℃) stirring at room temperature 2 hours, (1.03ml 8.65mmol) and with this mixture stirred under room temperature 18 hours to add bromotoluene., this mixture with EtOAc diluted, be cooled to 0 ℃ and add 10% aqueous citric acid solution and make acid (pH3), separate organic layer, water and salt water washing, dry (MgSO thereafter ₄), and under reduced pressure be concentrated into drying.Residual yellow oil chromatography purifying (SiO ₂, elutriant: hexane-ethyl acetate, 9: 1), can obtain 4(S)-benzyloxy-uncle 1-(-butoxy carbonyl) tetramethyleneimine-2(S)-carboxylic acid benzyl fat (301mg, 70%).

(c) (301mg 0.73mmol) is dissolved in MeOH/H will newly to get compound ₂O(2: 1,4ml) in, with this solution stirring and be cooled to 0 ℃.Add the aqueous sodium hydroxide solution (1.16ml) of 2M, after 10 minutes, make mixture be warming up to room temperature, under same temperature, stirred 18 hours then.Use Et thereafter, ₂The O/ hexane (1: 1,10ml) and water (5ml) reactant is diluted.Water phase separated is used Et ₂O/ hexane (1: 1) extraction 2 times is cooled to 0 ℃, and the aqueous citric acid solution with 10% makes acid (pH3), and extracts (3 times) with EtOAc.Blended EtOAc extract water (2 times) and salt water washing, dry (MgSO ₄) and under reduced pressure concentrate.Residue drying under high vacuum is obtained 4(S)-benzyloxy-uncle 1-(-butoxy carbonyl)-the quantitative product of tetramethyleneimine-2(S)-carboxylic acid.

(d) will be at CH ₂Cl ₂In the 0.2M of new compound

Solution (234.7ml, 0.73 mmol) is added to DIEA(127 μ l, 0.74m mol) in, add uncle-butylamine (84.4 μ l, 0.803m mol) and BOP(387mg subsequently, 0.0876m mol).By fixed time testing the pH value of reaction mixture is maintained 8(when needed, can add DIEA) time, at room temperature reaction mixture was stirred 3 hours.With EtOAc compound of reaction diluted, and use saturated NaHCO thereafter ₃The aqueous solution (2 times), water and salt solution in turn wash.With organic layer drying (MgSO ₄), and under reduced pressure be concentrated into drying.With the yellow oil of gained with quick chromatography (flash chromatography) (Sio ₂, elutriant: hexane-EtOAc, 7: 36: 4 afterwards) and purifying, can obtain title compound (252mg, 92%). ¹NMR（CDCl ₃）δ7.40-7.25（m，5H），6.05（broad s，1H），4.6-4.35（broad d，2H），4.2-4.05（m，2H），3.8-3.55（m，2H），2.55-2.1（m，2H），1.46（s，9H），1.20（broad s，9H）。

Example 2

1-3(S)-amino-2(R)-the hydroxy-4-phenyl butyl }-4(S)-benzyloxy-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (formula 4: R ¹=C(CH ₃) ₃, and Y-OCH ₂Ph; C(O) NHR ¹/ Y=cis) preparation

(a) will be in 6N hydrochloric acid/diox the solution of (250mg, the 0.664m mol) of the title compound of example 1, at room temperature stirred 20 minutes, under reduced pressure be concentrated into drying then.With residue EtOAc(10ml) dilute with the 2N NaOH aqueous solution (3ml).This mixture was at room temperature stirred 15 minutes.Separate organic layer, with the water and the salt water washing of minimum, dry (MgSO ₄) and under decompression, be concentrated into drying.Residue is dry under high vacuum, can get 4(S)-benzyloxy-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides, the i.e. carboxylic acid amides of formula 3 R wherein ¹Be C(CH ₃) ₃, Y is OCH ₂Ph{C(O) NHR ¹/ Y=cis }.

(b) will be newly compound at anhydrous EtOH(5ml) in 3(S)-(carbobenzoxy-(Cbz))-1,2(R)-epoxy-4-phenyl butane (180mg, 0.604m mol) mix, be exactly formula 2 epoxide, wherein X is Z, the people such as B.K.Handa that see above with mixture reflux 18 hours, under reduced pressure are concentrated into drying then.By quick chromatography with residue purifying (SiO ₂, elutriant: CHCl ₃-MeOH, 39: 1 19: 1 then), can get 4(S)-benzyloxy-1-3(S)-{ (carbobenzoxy-(Cbz)) amino }-2(R)-hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (220mg, 63%), be the white foam shape.

(c) the new compound (220mg, 0.384m mol) that gets can get title compound through hydrogenolytic cleavage (5%Pd/C, the normal atmosphere of 1 hydrogen, MeOH, 3.5 hours), and this compound is used immediately according to the coupling method of following example.

Example 3

4(S)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz)) valyl amino-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (formula 1: X=Z, B=Val, R ¹=C(CH ₃) ₃,

And Y=OCH ₂Ph; C(O) NHR ¹/ Y=cis) preparation

Prepare title compound according to following coupling method:

With DIEA(33.4 μ l, 0.192m mol), the amino acid Z-Val-OH(53.1mg of protection, 0.211m mol) and Bop(102mg, 0.23m mol) be added at CH ₂Cl ₂In the 0.2M solution of the title compound (0.192m mol) of middle example 2.When mixture is at room temperature stirred 2 hours, by fixed time testing the pH value of reaction mixture is maintained 8, and can add DIEA when needed., reaction mixture with EtOAc diluted, with saturated NaHCO thereafter ₃The aqueous solution (2 times), water and salt solution wash successively.With organic layer drying (MgSO ₄) and concentrated down in decompression.With residue with quick chromatography purifying (SiO ₂, elutriant: CHCl ₃-MeOH, 39: 1), can get the title compound of this example, be white solid (108mg, 83%).The FAB mass spectrum, m/z:673.3(M+H) ⁺

Example 4

4(R)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz)) valyl amino-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (formula 1: X=Z, B=Val, R ¹=C(CH ₃) ₃With

Y=OCH ₂Ph; C(O) NHR ¹/ Y=is trans) preparation

Method according to example 1,2,3 orders, but with example 1(a) 4(S of joint in the step)-hydroxyl pyrrolidine-2(S)-carboxylic acid is with the 4(R of equivalent)-hydroxyl pyrrolidine-2(S)-carboxylic acid (trans-4-oxyproline-2-carboxylic acid) substitutes, see above S.G.Ramaswamy and E.Adams then can obtain title compound.The FAB mass spectrum, m/z:673.3(M+H) ⁺

Example 5

4(R)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz)) and asparagyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (formula 1: X=Z, B=Asn, R ¹=C(CH ₃) ₃With

Y=OCH ₂Ph; C(O) NHR ¹/ Y=is trans) preparation

According to example 1, the method of 2 orders, but with the 4(S in (a) joint step of example 1)-hydroxyl pyrrolidine-2(S)-the carboxylic acid 4(R of equivalent)-hydroxyl pyrrolidine-2(S) carboxylic acid substitutes, and the 1-that will so obtain { 3(S)-amino-2(R)-hydroxy-4-phenyl butyl }-4(R)-benzyloxy-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amine carries out following coupling step, then can get title compound.

(20.1mg, 0.148m mol) is added to the THF(2mL that is cooled to 0 ℃ with I-hydroxybenzotriazole) in N, in N '-dicyclohexyl carbodiimide (34mg, the 0.165m mol) solution.This mixture was stirred 15 minutes.Will be at DMF(1mL) in through the amino acid Z-Asn-OH(395mg of protection; 0.148m mol) solution and aforesaid 1-{ 3(S)-amino-2(R)-hydroxy-4-phenyl butyl }-4(R)-benzyloxy-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (35.4mg, 0.083m mol) solution is added in this mixture.Make this mixture be warming up to room temperature slowly, stirred subsequently 18 hours.Thereafter, with EtOAc with this mixture diluted.Organic phase is separated, with saturated NaHCO ₃The aqueous solution, water and salt water washing, dry (MgSO ₄), under reduced pressure be concentrated into drying.With quick chromatography (SiO ₂, elutriant: EtOAc/MeOH, 97: 3 19: 1 then) and with this white residue purifying, can get the title compound of this example.The EI mass spectrum, m/e:689.2(M+2H) ⁺

(note: before exemplified at N, under the situation that N '-dicyclohexyl carbodiimide exists, utilize the coupling method of I-hydroxybenzotriazole to be represented as the preferably coupling method of preparation B for formula 1 compound of amino-acid residue Asn.）

Example 6

4(S)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz)) and asparagyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides (formula 1: X=Z, B=Asn, R ¹-C(CH ₃) ₃, and Y=OCH ₂Ph; C(O) NHR ¹/ Y=cis) preparation

According to example 1 and the method for 2 orders and the coupling method of example 5, can obtain title compound.The FAB mass spectrum, m/z:688.4(M+H) ⁺710.4(M+Na) ⁺

Example 7

1-{ 3-(S)-{ { N-(carbobenzoxy-(Cbz))-valyl } amino }-2(R)-hydroxy-4-phenyl butyl }-tetramethyleneimine of N-tert-butyl-4(S)-(2-methyl propoxy-)-2(S)-carboxylic acid amides { formula 1: X=Z, B=Val, R ¹=C(CH ₃) ₃And Y=OCH ₂CH(CH ₃) ₃; C(O) NHR ¹/ Y=cis } preparation

According to the method for example 1,2 and 3 orders, but the 2-methyl-propyl bromine of the bromotoluene in (b) joint step of example 1 with equivalent substituted, then can obtain title compound.The EI mass spectrum, m/z:583.3(MH ₂-C ₄H ₃) ⁺

Example 8

1-({ 3(S)-{ { N-(carbobenzoxy-(Cbz))-valyl } amino }-2(R)-hydroxy-4-phenyl butyl }-tetramethyleneimine of N-tert-butyl-4(R)-(2-methyl propoxy-)-2(S)-carboxylic acid amides { formula 1: X=Z, B=Val, R ¹=C(CH ₃) ₃And Y=OCH ₂CH(CH ₃) ₂; C(O) NHR ¹/ Y=is trans } preparation

Method according to example 1,2 and 3 orders, but with the 4(S in (a) joint step of example 1)-hydroxyl pyrrolidine-2(S)-the carboxylic acid 4(R of equivalent)-hydroxyl pyrrolidine-2(S)-carboxylic acid is alternative, at example 1(b) bromotoluene of joint in the step substitute with the 2-methyl-propyl bromine of equivalent, then can obtain title compound.The EI mass spectrum, m/e:583.3(MH ₂-C ₄H ₃) ⁺

Example 9

4(R)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz))-valyl amino-2(R)-the hydroxy-4-phenyl butyl }-N-cyclopropyl tetramethyleneimine-2(S)-carboxylic acid amides (formula 1: X=Z, B=Val, R ¹=cyclopropyl and Y=OCH ₂Ph; C(O) NHR ¹/ Y=is trans) preparation

Method according to example 1,2 and 3 orders, but with example 1(a) 4(S in the joint)-hydroxyl pyrrolidine-2(S)-carboxylic acid is with the 4(R of equivalent)-hydroxyl pyrrolidine-2(S)-carboxylic acid substitutes, and in example 1(d) uncle-butylamine in the joint substitutes with the cyclopropylamine of equivalent, then can obtain title compound.The EI mass spectrum, m/e:657.5(M+H) ⁺

Example 10

4-benzyl-1-{ 3(S)-{ { N-(carbobenzoxy-(Cbz))-asparagyl } amino }-2(R)-hydroxy-4-phenyl butyl }-4(R of N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides; 4(R) and (formula 1: X=Z of isomer 4(S) S); B=Asn, R ¹=C(CH ₃) ₃And Y=BzL) preparation

By with above-mentioned by F.Soucy, the described method of D.Wernic and P.Beaulieu, 4-benzyl-uncle 1-(-butoxy carbonyl) 4(R of tetramethyleneimine-2(S)-carboxylic acid) and 4(S) mixture of diastereomer is (3: 2, W/W), can obtain by Serine lactone and 3-phenyl-2-propylene bromine.According to example 1(d) joint step, under the situation that BOP exists, this diastereoisomeric mixture and uncle-butylamine coupling can obtain 4-benzyl-N-tert-butyl-uncle 1-(-butoxy carbonyl) 4(R of tetramethyleneimine-2(S)-carboxylic acid amides) and corresponding diastereoisomeric mixture of isomer 4(S).Thereafter, according to example 2(b) step of joint and use the carboxylic acid amides of new diastereoisomeric mixture as formula 3, then can obtain the 4(R of 4-benzyl-1-{ 3(S)-{ (carbobenzoxy-(Cbz))-amino }-2(R)-hydroxy-4-phenyl-butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides) and 4(S) corresponding diastereoisomeric mixture of isomer.The FAB mass spectrum, m/z:558(M+H) ⁺Then; coupling method according to example 5; the amino acid Z-Asn-OH of diastereoisomeric mixture of the latter and N-protected reaction can get 4-benzyl-1-{ 3(S)-{ { N-(carbobenzoxy-(Cbz)) asparagyl } amino }-2(R)-hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-4R of carboxylic acid amides and corresponding diastereoisomeric mixture of 4S isomer.The FAB mass spectrum, m/z:672(M+H) ⁺

With isolating these two isomer of HPLC technology, can obtain corresponding pure 4R and 4S isomer.Further illustrate.With 20mg above the sample dissolution of last-mentioned mixture in 50% aqueous acetic acid of 2.5mL (initial condition), be contained in Whatman Magnum 9

In, C ₁₀18 silicomethane pilums are (in 0.94 * 50cm).Initial column equilibration condition is as follows: 10%A and 90%B(pump A:0.06% trifluoracetic acid are in acetonitrile, and pump B:0.06% trifluoracetic acid is in water).In case the peak corresponding to acetic acid (in the solvent front) is passed through, a linear gradient is just followed.The isomer separation scheme is as follows: under 3mL/min and 230nm, and the A5 of 10-30% minute, 30% A10 minute, subsequently, the A110 of 30-100% minute.4(R) isomer and 4(S) isomer is respectively at 60%A(9.2mg) and 63% A(8.3mg) under be collected.

Example 11

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino }-butyl }-4(R)-(2-pyrimidine-based sulfur-base) tetramethyleneimine-2(S)-carboxylic acid amides (formula 1:X=2-quinoline carbonyl, B=Val, R ¹=(C(CH ₃) ₃With the Y=2-pyrimidine-based sulfur-base) preparation

(a) with the acid (17.5g, 75.6m mol are described in example 1(a) of N-protected joint) be dissolved in CH ₂Cl ₂(300mL) and EIDA(13mL, 76.6m mol).(8.73mL, 83.1m mol) is added in this solution with uncle-butylamine, adds BOP(40g subsequently, 90.7m mol) and DIEA(13mL, 151m mol).Mixture was stirred under room temperature 7 hours, dilute with EtOAc subsequently.Separate organic phase, with saturated NaHCO ₃The aqueous solution (2 times), water (2 times) and salt solution (2 times) washing, dry (MgSO ₄) and be evaporated to drying.Use Et ₂O/EtOAc(9: 1), be collected on the strainer, use Et with the solid residue development ₂The O washing is also dry, obtains N-tert-butyl-uncle 1-(-butoxy carbonyl)-4(R)-hydroxyl pyrrolidine-2(S)-carboxylic acid amides (15.6g, 72%).

(b) will be newly compound (5.0g, 17.5m mol) be dissolved in toluene/THF(3: 1,175mL) in.At room temperature triphenyl phosphine (5.72g, 21.8m mol) and imidazoles (1.08g, 30.5m mol) are added in this solution.This mixture is warming up to 45-50 ℃, adds iodine (5.54g, 21.8m mol) and under 45-50 ℃, firmly stirred the mixture 80 minutes.Thereafter, with reaction mixture cooling and use Et ₂O and water dilution.Separate organic layer, with saturated NaHCO ₃The aqueous solution (1 time) and salt solution (1 time) washing, dry (MgSO ₄) and be evaporated to drying, then can get brown oil, include a little solid (triphenyl phosphine oxide compound).With oily solid Et ₂O development and with solid collection on strainer.Filtrate is evaporated to drying, then can gets brown oil.With quick chromatography with this oily matter purifying (SiO ₂, elutriant: EtOAc/ hexane, 1: 4), can get N-tert-butyl-1-(tert-butyl carbonyl)-4(S)-iodol alkane-2(S)-carboxylic acid amides, be yellow solid (4.83g, 70%). ¹NMR（CDCl ₃）δ6.2-6.0（broads，1H），4.27-4.0（m，3H），3.75-3.55（m，1H），2.9-2.5（m，2H），1.47（s，9H），1.38（s，9H）。

(c) 2-pyrimidine mercaptan (1.06g, 9.46m mol) is added to refrigerative (0 ℃) DMF(10mL in batches) in sodium hydride (99%, 182mg, 7.57m mol) suspension in.Mixture was stirred 30 minutes under identical temperature.Thereafter, will be at DMF(5ml) in the solution of product (1.5g, 3.79m mol) of above (b) joint splash into this mixture.This reaction mixture after stirring 18 hours under the room temperature, is diluted with EtOAc and water again.Separate organic phase, the NaOH aqueous solution (2 times) of using cold water (1 time), 1N is at salt solution (1 time) washing, dry (MgSO ₄), and be evaporated to drying, obtain solid.With this solid Et ₂O development can get N-tert-butyl-uncle 1-(-butoxy carbonyl)-4(R)-(2-pyrimidine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides, be a kind of pale solid.

¹H NMR（CDCl ₃）δ8.53-8.51（d，J＝4.85Hz，2H），7.01-6.96（t，J-4.85，10.0Hz，1H），5.97-5.75（broad s，1H），4.4-4.2（m，2H），4.1-3.91（m，1H），3.70-3.35（m，2H），2.92-2.75（m，1H），1.47（s，9H），1.36（s，9H）。FAB mass spectrum (m/z): 381(M+H) ⁺, 403(M+Na) ⁺

(d) according to example 2(a) with (b) joint step; to newly get compound and go protection; and with the reaction of the epoxide (wherein X is Boc) of formula 2, can get the tetramethyleneimine of N-tert-butyl-1-{ 3(S)-{ (uncle-butoxy carbonyl) amino }-2(R)-hydroxy-4-phenyl butyl }-4(R)-(2-pyrimidine sulfenyl)-2(S)-carboxylic acid amides.The FAB mass spectrum, m/z; 544(M+H) ⁺, 566(M+Na) ⁺

Then; according to example 2(a) joint step; to newly get compound and go protection; and, can get N-tert-butyl-1-{ 3(S)-{ { uncle N-(-butoxy carbonyl) valyl } amino }-2(R)-hydroxy-4-phenyl butyl according to the method and the Boc-Val-OH coupling of example 3 }-4(R)-(2-pyrimidine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides.FAB mass spectrum (m/z): 643(M+H) ⁺665(M+Na) ⁺

(e) new compound (887mg, the 1.38m mol) solution that gets in 6N hydrochloric acid/diox (7mL) was stirred under room temperature 20 minutes.Under reduced pressure solvent is removed.A kind of is the residue of white solid under high vacuum dry 20 minutes, is removed to protect amine accordingly, is hydrochloride.To newly get salt and be dissolved in CH ₂Cl ₂(7mL) with DIEA(481 μ L, 2.76m mol) in.Quinaldinic acid (263mg, 1.52m mol) and BOP(732mg, 1.66m mol) add in the solution of this salt.By fixed time testing, the pH value of this mixture is maintained 8(can add DIEA when needed) time, this reaction mixture was at room temperature stirred 5 hours., reaction mixture with EtOAc diluted, and use saturated NaHCO successively thereafter ₃The aqueous solution (2 times), water (2 times) and salt water washing.With organic layer drying (MgSO ₄), and under reduced pressure be concentrated into drying.The colorless oil of gained is with quick chromatography (SiO ₂, elutriant: hexane-EtOAc, 3: 71: 9 then) and purifying, get title compound, be white foam shape thing (750mg, 78%).With this foam Et ₂The O development can get title compound, is white solid (378mg, 40%).FAB mass spectrum (m/z:698(M+H) ⁺, 720(M+Na) ⁺The NMR of compound is consistent with specified structure.

Method according to this example; but the 2-pyrimidine mercaptan in (c) joint is substituted with 3-pyridine thiomethyl alcohol, then can get N-tert-butyl-1-(2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl }-amino butyl-4(R)-{ (3-picolyl) sulfenyl } tetramethyleneimine-2(S)-carboxylic acid amides.

FAB mass spectrum m/z:711(M+H) ⁺, 733(M+Na) ⁺

Once more; method according to this example; but the 2-pyrimidine mercaptan that (c) joint is interior is with 2; 6-dimethyl-4-hydroxy pyrimidine replaces; then can get N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino } butyl }-4(R)-(2,6-dimethyl-4-pyrimidyl } the oxygen base } tetramethyleneimine-2(S)-carboxylic acid amides.The FAB mass spectrum,

m/z：710（M+H），586（M+H-C ₆H N ₂O） ⁺。

Example 12

N-tert-butyl-1-3(S)-{ { 2; the 6-dimethyl phenoxy) ethanoyl } amino }-2(R)-hydroxy-4-phenyl-butyl }-4(R)-(2-pyrimidine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides (formula 1:X=(2; the 6-dimethyl phenoxy) ethanoyl, B is not for existing R ¹=C(CH ₃) ₃With Y be 2-pyrimidine sulfenyl) preparation

By remove the Boc protecting group with usual method; will be at example 11(d) tetramethyleneimine of N-tert-butyl-1-of describing in the joint { 3(S)-{ N-(tert-butyl carbonyl) amino }-2(R)-hydroxy-4-phenyl butyl }-4(R)-(2-pyrimidine sulfenyl)-2(S)-carboxylic acid amides; convert its corresponding primary amine to, i.e. uncle N--Ding-1-(3(S)-amino-2(R)-hydroxy-4-phenyl butyl)-4(R)-(2-pyrimidine sulfenyl) tetramethyleneimine-2-carboxylic acid amides.According to the method for example 3, will newly get compound and can get title compound with (2, the 6-dimethyl phenoxy) acetate coupling.The FAB mass spectrum, (m/Z:606(M+H) ⁺, 628(M+Na) ⁺

Method according to this example; but this primary amine is replaced with corresponding primary amine; be N-tert-butyl-1-(3(S)-amino-2(R)-hydroxy-4-phenyl butyl)-4(R)-{ (3-picolyl)-sulfenyl }-tetramethyleneimine-2-carboxylic acid amides (as N-tert-butyl-1-in the example 11 { 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino }-butyl }-4(R)-{ (3-picolyl) sulfenyl } intermediate of tetramethyleneimine-2(S)-carboxylic acid amides); obtain N-tert-butyl-1-3(S)-{ { (2; the 6-dimethyl phenoxy) ethanoyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-4(R)-{ (3-pyrimidine methyl)-sulfenyl } tetramethyleneimine-2(S)-carboxylic acid amides, FAB mass spectrum m/z:619(M+H) ⁺, 641(M+Na) ⁺

Example 13

Recombinant HIV albumen acyl HPLC analyzes

Enzyme: in intestinal bacteria, can give expression to hiv protease { construct pBRTlprt ⁺, see people such as W.G.Farmerie, Science, 236,305(1987), according to following conditions:

Except as otherwise noted, all solution are the aqueous solution.

(ⅰ) fermentation

Contain pBRTl prt ⁺The Bacillus coli cells of plasmid is used for inoculating the seed culture medium of being made up of the Lu-Bai meat soup (Luria-Bertani Broth) of the penbritin that contains 100 μ g/mL.Flask was cultivated 17 hours under vibrations in 37 ℃.The preparation flask that contains aseptic M9 meat soup replenishes the penbritin of 100 μ g/mL, with above-mentioned seed culture medium, with 1%(V/V) concentration inoculate.At the cumulative volume of each preparation in flask is 500mL in Ou Lunmaiershi (Erlenmeyer) flask of 2L.It is 0.6(λ=540 μ m that flask cultivation under 37 ℃ of earthquakes is reached corresponding optical density(OD) until cell concn) (not dilution).This timed interval is generally 3-4 hour.Then to flask replenish 5mM triisopropyl thiogalactoside (IPTG, Organics, Cleveland, Ohio USA), and continues to be cultured to cell concn and dilutes doubly down in 16-that optical density(OD) is 0.2.The phenyl methyl sulfonic acid fluoride (PMSF) that replenishes 1mM then is to flask, and is quickly cooled to 4 ℃.Bacterial cell is in 4 ℃ of centrifugal recovery down.Wet small pieces are stored down at-70 ℃.

(ⅱ) extraction of AG enzyme and preparation

Below all step all carry out down in 4 ℃, except as otherwise noted, mix thaw cell and buffer A three (hydroxymethyl) ethylamine hydrochloric acid of 50mM (Tris-HCl, pH7.4); 0.6mM ethylenediamine tetraacetic acid (EDTA) (EDTA); 0.375M NaCl, 0.2% Nonidet p

(BDH Chemical, Ltd., Povle, UK); The PMSF} of 1mM, its ratio is that 1 part of cell is to 9 parts of buffer A.(Celite 545 to add diatomite

, John Manville, Lompoc, CA, USA), its ratio be 2 parts to 1 part of wet cell weight.The mashed prod that produces at a high speed (about 20,000rpm) under at Waring

Carry out homogenizing 8 * 15 pulse per second (PPS)s on the industrial mixer.Cell debris/Celite

From centrifugal collection, use above-mentioned homogenization step that the small pieces that produce are extracted 1 part of wet solid with 4.5 parts buffer A.The supernatant liquor that mixing is come by above-mentioned two homogenization steps, and add solid (NH ₄) ₂SO ₄, with the protein precipitation of solubility, to produce 75% saturated final concentration.With this mixture vibrations 60 minutes and centrifugal collecting precipitate.The small pieces that produce are suspended in buffer B { 50mM Tris-HCl, pH8; 30mM NaCl, 1mM DL-dithiothreitol (DTT) (DTT); 1mM EDTA; 1mM PMSF; 10% glycerine } in, and to same damping fluid dialysis 18 hours.

With a part contain the proteinic dialysis extract of 150mg pack into bed be of a size of on Sephadex A 25 anion-exchange columns of long 70cm, diameter 2.5cm (Pharmacia, Uppsala, Sweden).With buffer B with 10cm/ hour linear rate of flow with sample elution equably.Mix and contain hiv protease active part (analysis of stating is as follows described), and by adding saturated (NH ₄) ₂SO ₄The aqueous solution is with the soluble protein precipitation, to produce 85% saturated total (NH ₄) ₂SO ₄Concentration is removed sedimentary protein with centrifugation, and with the small pieces that produce be dissolved in damping fluid C the 2-(4-morpholino of 50mM) ethyl sulfonic acid (MES), pH5.5; 150mM NaCl; 1mM DTT; 1mM EDTA; 10% glycerine }.This prepared product is chilled in-70 ℃ again to damping fluid C dialysis 18 hours.All crude extracts are all to contain the proteinic aliquots containig of the 150mg same procedure purifying of chromatographic analysis as the aforementioned.The whole prepared product of each batch is compiled, be distributed into the aliquots containig of 34 μ L and be stored in-70 ℃.Usually the whole protein that is reclaimed by the 20L fermented product is 300mg, and have 18.2m mol substrate hydrolysis/minute/the hiv protease specific activity of mg.

Before using, be the enzyme working solution with the 1/18(that buffering liquid is diluted to original concentration with these aliquots containigs), see below.

Substrate: VSFNFPQITL-NH ₂, MW 1164, see people such as Krausslich, Proc.Natl.Acad.Sci.USA, and 86,807(1989), as substrate.Substrate is made 10mM storage liquid in DMSO, be stored under 4 ℃.Before use, will store liquid and get 400 μ m solution (being the substrate working solution) with the damping fluid dilution.

Damping fluid: with MES(100mM), KCl(300mM) and EDTA(5mL) be dissolved in distilled water (90mL), and solution is adjusted to pH5.5 with the dense NaOH aqueous solution.To newly get solution and be diluted to 100mL to form damping fluid with water.

Step: (1) prepares analysis of mixtures by the substrate working solution that mixes 20 μ L, the solution of the test compound of 10 μ L in 10%DMSO and the enzyme working solution of 10 μ L.(2) analysis of mixtures was cultivated 30 minutes down in 37 ℃.(3) make the reaction all standing by the trifluoroacetic acid aqueous solution that adds 200 μ L2%.(4) utilize one to have step by step that gradient is Perkin-Elmer 3 * 3CRC8 post (Perkin Elmer Inc. of 4mL/min flow velocity, Norwalk, CT, USA), making the analysis of mixtures of 100 μ L all standings make substrate and product with HPLC (is VSFNF and PQITL-NH ₂) separate.

This gradient is as follows:

0.0-0.5 minute, 70%A/30%B;

0.5-3.0 minute, 67%A/33%B;

3.0-5.0 minute, 20%A/80%B;

5.0-6.5 minute, 70%A/30%B;

Wherein A is 3mM dodecyl sodium sulfate/0.05%H ₃PO ₄Yu Shuizhong, and B is 0.05%H ₃PO ₄In acetonitrile, when 210mM, monitor elution.

(5) with an object of reference, it is the analysis of mixtures of no test compound, stands step 2～4 simultaneously.

Suppress research: hydrolysate and residual former substrate are with peak heights or come quantitative with the integration at suitable HPLC peak.Use following relationship to calculate substrate conversion:

% conversion=(peak height of product or territory, peak summation)/(peak height of substrate and product or territory, peak summation) * 100

Being calculated as follows that the enzyme of test compound suppresses is described:

% inhibition=100-(% of analysis of mixtures transforms)/(% of object of reference transforms) * 100

Cause the concentration of 50% test compound that suppresses of HIV-proteolytic enzyme, i.e. IC ₅₀, can determine by following: determine the inhibition per-cent of enzyme, as the minimum value of three kinds of different concns of test compound.In view of the above, by the inhibition per-cent of enzyme is mapped to the concentration of test compound, determine IC with graphics ₅₀

IC as the example of formula 1 compound ₅₀, determine by Recombinant HIV proteolytic enzyme HPLC analysis, in being listed in the table below.

The table I

Table I (continuing)

Example 13

The following condition that is used for the antiviral effect of screening type 1 compound is borrowed in the plaque analysis of before having been delivered by people such as above-mentioned Harada with the HTLV-1 transformant.Why using the HTLV-1 transformant, is because the rapidity that HIV duplicates in this cell.

1. test compound is dissolved in the dimethyl sulfoxide (DMSO) to its concentration be 5mg/mL.The solution of gained can be stored in 4 ℃ to using.

2. (MA USA) is diluted to 4 times (4X) for Gibco Laboratories, St.Lawrence, the test final concentration in RPMI 1640 with the solution that obtains.In case dilution in RPMI 1640, this solution needed to be used for cell culture assays in 4 hours.

3. these 4 times of solution (50 μ L) are added in the triplicate aperture of 96 aperture flat-bottom microtiter plates, RPML(50 μ L) also be added in the contrast aperture.

4. will be at the HEPES-buffered RPMI of 50 μ L 1640(pH=7.2) in C8166 cell (5 * 10 ⁴), the foetal calf serum (FCS) of 10% heat inactivation, the gentamicin (perfect medium) of 12.5 μ L/mL be added in all apertures.

5. will be in the perfect medium of 100 μ L 50 times of TCID of H9/HTLV-III B storage liquid (being stored in the liquid nitrogen) as cell culture supernatant in 50%FCS ₅₀, be added in all apertures.The infectivity of virus storage liquid is tired and is determined by the terminal point dilution of C8166 cell as previous.Storage liquid is tired be stored in and can be stablized 6-12 month under-193 °.

6. then microtiter plate was put 72 hours on 37 ℃, the layer frame of the CO2gas incubator of 5% humidity.7. then flat board is taken out, and calculate plasmodial middle calculation in each aperture with low power phase contrast opticmicroscope.The every bunch of cell that shows the evidence that any synplasm forms is promptly counted a synplasm center.The contrast aperture should make every aperture have 25-75 synplasm center.

8. the inhibition per-cent of synplasm formation calculates with following formula:

% suppresses=100 * ((calculation in the synplasm in the contrast aperture)-(calculation in the synplasm in the test aperture))/((calculation in the synplasm in the contrast aperture))

Cause suppress that 50% synplasm forms the concentration of test compound, i.e. EC ₅₀, determine by employed technology of the serial dilution of working solution in step 3 and the figure that the inhibition per-cent that forms with the synplasm that observes is marked and drawed the different concns of test compound.

Under tabulate in the II, as the example of the compound of formula 1, the analytical results of listing is from the plaque analysis of this example.

The table II

Other compound of formula 1 is:

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino } butyl }-4(R)-(benzene sulfonyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino } butyl }-4(R)-(2-pyridine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino } butyl }-4(R)-(4-pyridine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino } butyl }-4(R)-(4,6-dimethyl-2-pyrimidine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-2(R)-hydroxyl-4-benzyl-3(S)-N-(2-pyridine carbonyl) valyl amino butyl-4(R)-benzene oxygen tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-2(R)-hydroxy-4-phenyl-3(S)-N-(2-pyridine carbonyl) and asparagyl } amino } butyl }-4(R)-benzene oxygen tetramethyleneimine-2(S)-carboxylic acid amides

N-cyclopentyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) leucyl } amino } butyl }-4(R)-(benzenesulfonyl) tetramethyleneimine-2(S)-carboxylic acid amides

1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) valyl } amino } butyl }-the N-(1-methylethyl)-4(R)-(2-pyridine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-cyclopropyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) asparagyl } amino } butyl }-4(R)-(4-pyridine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-quinoline carbonyl) isoleucyl } amino } butyl }-4(R)-(2-pyrimidine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-naphthalene carbonyl) valyl } amino } butyl }-4(R)-(4,6-dimethyl-2-pyrimidine sulfenyl) tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-pyridine carbonyl) isoleucyl } amino } butyl }-4(R) benzene oxygen tetramethyleneimine-2(S)-carboxylic acid amides

N-tert-butyl-1-{ 2(R)-hydroxy-4-phenyl-3(S)-{ { N-(2-pyridine carbonyl) asparagyl } amino } butyl }-4(R)-(thiophenyl) tetramethyleneimine-2(S)-carboxylic acid amides

Claims (10)

1, acceptable acid addition salt is gone up in a kind of compound of formula 1 or its treatment:

Wherein

X is R ²OC (O), R ²C (O) or R ²NR ²C (O), wherein R ²For

(i) low-carbon alkyl,

(ii) low-carbon naphthenic,

(iii) phenyl or with the mono-substituted phenyl of halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy,

(iv) phenyl (low-carbon (LC)) alkyl or phenyl (low-carbon (LC)) alkyl, wherein its aromatic base partly replaces with halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy list,

(v) 1-naphthyl or 2-naphthyl

(vi) (Het) or (Het)-(low-carbon alkyl), wherein Het represent five or hexa-atomic, contain 1 or 2 unit price heterocyclic radical that is selected from the heterocyclic atom of nitrogen, oxygen and sulphur, or

(vii) 2-quinolyl or 3-quinolyl,

R ³Be hydrogen or low-carbon alkyl;

Or X is R ^2AOCH ₂C (O), wherein R ^2AReplace two replacements or trisubstd phenyl for phenyl or with low-carbon alkyl or halogen list;

B is not for existing or divalent radical-NHCHR ⁴C (O)-, R wherein ⁴

Be low-carbon alkyl; Or low-carbon naphthenic; (low-carbon naphthenic)-(low-carbon alkyl) phenyl methyl; Or with hydroxyl, carboxyl, low-carbon (LC) carbalkoxy, aminocarboxyl, the mono-substituted low-carbon alkyl of (low-carbon alkyl) aminocarboxyl or two (low-carbon alkyl) aminocarboxyl;

R ¹Be low-carbon alkyl or low-carbon naphthenic;

Y is a low-carbon alkyl; Low-carbon naphthenic, phenyl or with the mono-substituted phenyl of halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy; Phenyl methyl or use halogen, hydroxyl, the mono-substituted phenyl methyl of low-carbon alkyl or low-carbon alkoxy; Or

Y is W (CH ₂) _nZ, wherein W be oxo, sulfo-, sulfinyl or

Sulfonyl, Z are low-carbon alkyl, phenyl or with the mono-substituted phenyl of halogen, hydroxyl, low-carbon alkyl or low-carbon alkoxy; Or (Het) wherein (Het) is described as defined above; With n be 0 or 1.

2, the compound of claim 1, or its acceptable acid addition salt in treatment, wherein X is R ²OC(O) or R ²C(O), R wherein ²Be low-carbon alkyl; Phenyl (low-carbon (LC)) alkyl; Phenyl (low-carbon (LC)) alkyl, wherein the position 4 usefulness chlorine of phenyl moiety, fluorine, hydroxyl, methyl or methoxy replace; The 1-naphthyl; The 2-naphthyl; The 2-furyl; The 2-thienyl; The 2-pyridyl; The 4-pyridyl; The 2-pyridylmethyl; 4-thiazolyl methyl or 2-quinolyl; Or X is R ^2AOCH ₂C(O) R wherein ^2AFor phenyl or position 2,

On 4 and 6, with low-carbon alkyl or halogen is single, double or trisubstd phenyl;

B is not for existing or being divalent radical-NHCHR ⁴C(O)-, R wherein ⁴Be low-carbon alkyl; Or with hydroxyl, low-carbon (LC) carbalkoxy, aminocarboxyl, the mono-substituted low-carbon alkyl of (low-carbon alkyl) aminocarboxyl or two (low-carbon alkyl) aminocarboxyl.

R ¹Be the 1-methylethyl, 1,1-dimethyl ethyl, 2-methyl-propyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

Y is a low-carbon naphthenic, phenyl, 4-chloro-phenyl-, 4-bromophenyl, 4-fluorophenyl, 4-aminomethyl phenyl, 4-methoxyphenyl, phenmethyl, (4-fluorophenyl) methyl or (4-aminomethyl phenyl) methyl; Or

Y is W(CH ₂)

Z, in the formula definition of W and n as above, Z is a low-carbon alkyl, phenyl, 2-furyl, 2-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 4-thiazolyl, 2-pyrimidyl, 4,6-dimethyl-2-pyrimidyl or 2,6-dimethyl-4-pyrimidyl.

3, acceptable acid addition salt is gone up in the compound of claim 2 or its treatment, and wherein X is uncle-butoxy carbonyl, carbobenzoxy-(Cbz), (4-chloro-phenyl-) methoxycarbonyl, (4-hydroxyphenyl) methoxycarbonyl, (4-methoxyphenyl) methoxycarbonyl, ethanoyl, benzoyl, 1-naphthalene carbonyl, 2-naphthalene carbonyl, 2-pyridine methoxycarbonyl, or 2-quinoline carbonyl, benzene oxygen ethanoyl, (2-methylphenoxy) ethanoyl, (2, the 4-dimethyl phenoxy) ethanoyl, (2, the 6-dimethyl phenoxy) ethanoyl, (2,4,6-trimethylammonium phenoxy group) ethanoyl, (4-chlorophenoxy) ethanoyl or (4-fluoro-2,6-dimethyl phenoxy) ethanoyl;

B is not for existing or being divalent radical-NHCHR ⁴C(O)-, R in the formula ⁴Be the 1-methylethyl, 1,1-dimethyl ethyl, 1-methyl-propyl, 2-methyl-propyl, methoxycarbonyl methyl, ethoxycarbonyl-methyl or amino carbonyl methyl;

R ¹Be 1,1-dimethyl ethyl or cyclopropyl;

Y is a cyclohexyl, phenyl, 4-chloro-phenyl-; the 4-fluorophenyl, 4-methoxyphenyl, benzyl; (4-methoxyphenyl) methyl, 2-methyl propoxy-, phenoxy group; the 2-pyridyloxy, 3-pyridyloxy, 4-pyridyloxy; the 2-2-pyrimidinyl oxy, 4,6-dimethyl-2-2-pyrimidinyl oxy; 2,6-dimethyl-4-2-pyrimidinyl oxy, benzyloxy; 2-pyridine methoxyl group, 4-thiazolyl methoxyl group, 2-pyrimidyl methoxyl group; thiophenyl; the benzene sulfinyl; the benzene sulfonyl; 2-pyridyl sulfenyl; 3-pyridyl sulfenyl; 4-pyridyl sulfenyl, 2-pyrimidine-based sulfur-base, 4; 6-dimethyl sulfenyl-2-pyrimidine sulfenyl; benzylthio-, benzyl sulfinyl, benzyl sulfonyl; (2-picolyl) sulfenyl, (3-picolyl) sulfenyl or (4-picolyl) sulfenyl.

4, acceptable acid addition salt is gone up in the compound of claim 3 or its treatment, and wherein X is uncle-butoxy carbonyl, carbobenzoxy-(Cbz), ethanoyl, 2-naphthyl carbonyl, 2-pyridyl methoxycarbonyl, 2-quinolyl carbonyl;

B is valyl, isoleucyl or asparagyl;

R ¹Be 1,1-dimethyl ethyl or cyclopropyl; With

Y is a phenyl, benzyl, phenoxy group, 2-2-pyrimidinyl oxy, 2; 6-dimethyl-4-2-pyrimidinyl oxy, benzyloxy, thiophenyl, benzene sulfonyl, 2-pyridine sulfenyl; 3-pyridine sulfenyl, 4-pyridine sulfenyl, 2-pyrimidine sulfenyl, 4,6-dimethyl-2-pyrimidine sulfenyl or (3-picolyl) sulfenyl.

5, the compound of claim 1, or the last acceptable acid addition salt of its treatment, wherein X is (2-methylphenoxy) ethanoyl, (2, the 4-dimethyl phenoxy) ethanoyl, (2, the 6-dimethyl phenoxy) ethanoyl or (2,4,6-trimethylammonium phenoxy group) ethanoyl; B is not for existing; R ¹Be 1, the 1-dimethyl ethyl; And Y is a phenyl, benzyl, phenoxy group, 2-2-pyrimidinyl oxy, 2; 6-dimethyl-4-2-pyrimidinyl oxy, benzyloxy, thiophenyl, benzene sulfonyl, 2-pyridine sulfenyl; 3-pyridine sulfenyl, 4-pyridine sulfenyl, 2-pyrimidine sulfenyl, 4,6-dimethyl-2-pyrimidine sulfenyl or (3-picolyl) sulfenyl.

6, the compound of claim 1 is selected from: 4(S)-benzyloxy-1-3-(S)-the N-(carbobenzoxy-(Cbz)) valyl amino-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides,

4(R)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz)) valyl amino-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides,

4(R)-benzyloxy-1-3-(S)-the N-(carbobenzoxy-(Cbz)) and asparagyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides,

4(S)-benzyloxy-1-3-(S)-the N-(carbobenzoxy-(Cbz)) and asparagyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides,

1-{ 3-(S)-{ { N-(carbobenzoxy-(Cbz)) valyl } amino }-2(R)-hydroxy-4-phenyl butyl }-tetramethyleneimine of N-tert-butyl-4(S)-(2-methyl propoxy-)-2(S)-carboxylic acid amides,

1-{ 3(S)-{ { N-(carbobenzoxy-(Cbz)) valyl } amino }-2(R)-hydroxy-4-phenyl butyl }-tetramethyleneimine of N-tert-butyl-4(R)-(2-methyl propoxy-)-2(S)-carboxylic acid amides,

4(R)-benzyloxy-1-3(S)-the N-(carbobenzoxy-(Cbz)) valyl amino-2(R)-the hydroxy-4-phenyl butyl }-N-cyclopropyl tetramethyleneimine-2(S)-carboxylic acid amides,

4(R)-benzyl-1-3(S)-the N-(carbobenzoxy-(Cbz)) and asparagyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides,

4(S)-benzyl-1-3(S)-the N-(carbobenzoxy-(Cbz)) and asparagyl } amino }-2(R)-the hydroxy-4-phenyl butyl }-N-tert-butyl tetramethyleneimine-2(S)-carboxylic acid amides,

N-tert-butyl-1-2-(R)-hydroxy-4-phenyl-3(S)-N-(2-quinoline carbonyl) valyl amino butyl-4(R)-(2-pyrimidine sulfenyl)-tetramethyleneimine-2(S)-carboxylic acid amides,

N-tert-butyl-1-2(R)-hydroxy-4-phenyl-3(S)-N-(2-quinoline carbonyl) valyl amino butyl-4(R)-and (3-picolyl)-sulfenyl } tetramethyleneimine-2(S)-carboxylic acid amides,

N-tert-butyl-1-2-(R)-hydroxy-4-phenyl-3(S)-N-(2-quinoline carbonyl) valyl amino butyl-4(R)-(2,6-dimethyl-4-pyrimidyl } the oxygen base } tetramethyleneimine-2(S)-carboxylic acid amides,

N-tert-butyl-1-{ 3(S)-{ { 2, the 6-dimethyl phenoxy) ethanoyl } amino }-2(R)-hydroxy-4-phenyl butyl }-4(R)-(2-pyrimidine sulfenyl)-tetramethyleneimine-2(S)-carboxylic acid amides and

N-tert-butyl-1-{ 3(S)-{ { 2, the 6-dimethyl phenoxy) ethanoyl } amino }-2(R)-hydroxy-4-phenyl butyl }-4(R)-{ (3-picolyl)-sulfenyl } tetramethyleneimine-2(S)-carboxylic acid amides.

7, a kind of pharmaceutical composition comprises the compound of claim 1, or acceptable salt and pharmaceutically useful carrier are gone up in its treatment.

8, a kind ofly treat the method that human body HIV infects, comprise the compound of the claim 1 of taking significant quantity, or acceptable salt is gone up in its treatment.

9, a kind of method that is used to protect human body cell opposing HIV morbidity comprises with compound or its treatment of the claim 1 of anti-HIV significant quantity and goes up acceptable salt, handles this cell.

10, a kind of compound of claim 1 or method that acceptable acid addition salt is gone up in its treatment of preparing comprises:

(a) epoxide of wushu 2

Wherein X as defined in claim 1, with the tetramethyleneimine carboxylic acid amides reaction of formula 3,

R wherein ¹Defined with Y such as claim 1, can obtain the respective compound of formula 1, X wherein, R ¹With defined in Y such as this claim, and B is not for existing; Or

R wherein ¹With defined in Y such as this claim, with the activity derivatives reaction of carboxylic acid X-OH, wherein X is R ²C(O) or R ^2AOCH ₂(O), it defines with claim 1, can obtain the respective compound of formula 1, and wherein X is R ²C(O) or R ^2AOCH ₂C(O), it defines with this claim, R ¹With Y as defining in this claim, and B is not for existing; Or

(c) with the compound of formula 4, R wherein ¹With defined in Y such as this claim, under the situation that coupling agent exists, with formula X-NHCHR ⁴The a-amino acid coupling of COOH, wherein X is R ⁴Defined as claim 1, can obtain the respective compound of formula 1, X wherein, R ¹With defined in Y such as this claim, and B is divalent radical-NHCHR ⁴C(O)-, R wherein ⁴As defined in this claim; Or

(d) with the compound of formula 5

R wherein ¹, R ⁴With defined in Y such as this claim, with carboxylic acid X-OH activity derivatives reaction, wherein X is R ²C(O) or R ^2AOCH ₂C(O), as defined in this claim, can obtain the respective compound of formula 1, wherein X is R ²C(O) or R ^2AOCH ₂C(O), as defined in this claim, R ¹With defined in Y such as this claim, and B is divalent radical-NHCHR ⁴C(O)-, R wherein ⁴As defined in this claim; With

(e) if want, with aforementioned (a), (b); (c) or (d) compound of the formula 1 that is obtained in the joint is transformed into acceptable acid addition salt on the corresponding treatment.