WO2003006060A1 - Sh3 domain binding inhibitors - Google Patents

Abstract

SH3 domain binding inhibitors containing, as the active ingredient, nonpeptidic compounds having an activity of inhibiting SH3 domain binding, low-molecular weight compounds having an activity of inhibiting SH3 domain binding and a molecular weight of less than 750, in particular, compounds represented by the above general formulae (I) and (II), cytochalasins or pharmacologically acceptable salts thereof; and compounds represented by the above general formulae (Va), (Vb) and (VI) and pharmacologically acceptable salts thereof.

Description

 Specification

 SH3 domain binding inhibitor

 SH3 domain binding inhibitor

The present invention relates to an SH3 domain binding inhibitor containing a non-peptide compound or a pharmacologically acceptable salt thereof as an active ingredient. The present invention also relates to compounds useful as SH3 domain binding inhibitors. Various receptors exist on the cell membrane and are activated in response to extracellular stimuli to transmit their signals into cells. Various signal molecules gather on the cytoplasmic side of the receptor to form a complex. Depending on the type of stimulus, each signal molecule elicits a different physiological effect. The domain structure plays an important role in the formation of signal molecule complexes. Among them, the Src homology domain 3 (SH3 domain) was discovered as a region of high homology between the Src family — a domain consisting of approximately 60 amino acids, located in various proteins, and containing a proline-containing sequence (proline-rich). (Science, 252, 668 (1991), 257, 803 (1992), Faseb's Journal (FASEB J.), Vol. 14, p. 231 (2000)]. Known proteins having a proline-rich sequence include HIV-1Nef, p22-phox, p47-phox, Sam68, Sosl, Zol, Dynamic c-Cbl and the like [for example, EMBO J., 14 vol. , 484 (1995), Proceding of the National 'Academy' of Science ',' _Ob 'The' _U ',' S 'A ( _proc . Natl. Acad. Sci. USA), Vol. 91, p. 10650 (1994), J. Immunol. (J. Immunol.), Vol. 157, p. 1226 (1996), J. Immunol. (J. Immunol.), J. Immunol. Biol. Chem.), 270, 9115 (1995), etc.]. Proteins having an SH3 domain have been found in, for example, all eukaryotes, viruses such as HIV, and the like, and are considered to have universal functions. For example, Frc, which is a Src family kinase, Ras-GAP, PLC, PI3K, Abl, Btk, Lyn, Hck, Fgr, Proteins with enzyme activity such as Yes, adapter proteins without enzyme activity such as Grb2, Neks Vav, etc., p40-phox, p47-phox, p67-phox, etc. which are components of the NADPH oxidase complex [Proceding Prop. Natl. Acad. Sci. USA, Vol. 91, p. 10650 (1994) It is known that Txk, Tec, Tsk, Crk, Cortactin and the like have an SH3 domain. It is thought that protein-protein binding via the SH3 domain plays a role in forming a protein complex through an interaction with an appropriate affinity.

 Protein-protein binding via the SH3 domain exists between signal molecules that are thought to be a cause in various diseases such as cancer, AIDS (acquired immune deficiency syndrome), allergy, etc. [Biopolymer. 383 pages (1997)]. This suggests that SH3 domain binding inhibitors that inhibit protein-protein binding via the SH3 domain (SH3 domain binding inhibitory activity) are considered to be effective as therapeutic agents for these diseases, such as in vivo carcinogenesis. It can be used for many applications including the regulation of disease-causing proteins that involve E. coli.

In the peptide, the mutation in the SH3 domain binding sequence [Cell, Vol. 76, p. 933 (1994), Chemical 'Biology (Chem. Biol.), Vol. 7, R3-R8 (2000 And screening of peptides that bind to the SH3 domain of various SH3 domain proteins such as Src and PI3K ヽ Abl from combinatorial phage libraries (W095 / 24419, JP 2000-506522) Also, a peptide (WO96 / 21011) that specifically binds to the SH3 domain of c-Crk has been reported. Among peptide-modified peptide compounds, spirolactam compounds that inhibit SH3 domain binding (WO98 / 54208), and biases in which synthetic peptides containing an amino acid sequence having a proline skeleton in their structures are immobilized. [Journal of Op. American Chemical 'Society (J. Am. Chem. Soc.), Vol. 118, p. 287 (1996)]. The smallest of these compounds has a molecular weight of 790, but has a value of 220 / mol / L, The cohesive strength was weak [Journal 'Op' American 'Chemical' Society (J. Am. Chem. Soc.), Vol. 118, p. 287 (1996)].

 The SH3 domain binding inhibitors reported so far have been peptides or peptide compounds based on hydrophobic amino acids such as proline with a molecular weight of 750 or more. Peptides or peptidic compounds, for example, generally have poor blood stability, poor oral absorption, high molecular weight and are not easily incorporated into cells, and are easy to manufacture due to their complex structure However, it is costly, and it has many difficulties to use it as a therapeutic agent for diseases associated with SH3 domain binding.

 UCS15A (Luminacin C2 / SP4228A) is published in The Journal of Antibiotics, 53, 579 (2000), No. 58-116686, and No. 63-22583. No. 61-293920, No. 62-294619, No. 63-48213, No. 8-268888, Bone resorption inhibitory action, bactericidal action, immunosuppressive action, anti-trichophyton action, anti-tumor It is described as an active compound.

 Cytochalasins (cytochalasin), oral sericaracin (Rosellichalasiii), epoxy cytochalasin (epoxycytochalasii ^, force etoglobosin, chanoglobosin, penochalasin, and blood vessels such as aspochalasin) Biological activities such as an inhibitory effect on renewal (WO98 / 41205) are known.

 昍 Indication

 An object of the present invention is to provide an SH3 domain binding inhibitor containing a non-peptide compound or a pharmacologically acceptable salt thereof as an active ingredient. Another object of the present invention is to provide a compound useful as an SH3 domain binding inhibitor or a pharmacologically acceptable salt thereof.

The present invention relates to the following [1] to [ ⁵¹ ].

 [1] An SH3 domain binding inhibitor comprising, as an active ingredient, a non-peptide compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof.

[2] The SH3 domain binding inhibitor according to the above [1], wherein the non-peptide compound is a low-molecular compound having a molecular weight of less than 750. [3] The non-peptidic compound having SH3 domain binding inhibitory activity is represented by the general formula (I)

(Wherein ^,, R ³ \ R ^3b and R ⁴ are the same or different, and each represents a hydrogen atom, hydroxy, carboxy, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkoxy, or substituted or unsubstituted lower alkoxy. R ^3a and R ^3b together represent an oxygen atom; R ²³ and R ^2b are the same or different and are a hydrogen atom, substituted or unsubstituted lower alkyl or substituted or unsubstituted R ^5a and R ^5a and are the same or different and are ^{independently} a hydrogen atom, hydroxy, carboxy, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkoxy, substituted or unsubstituted lower alkenyl, substituted or Unsubstituted lower alkanoyloxy, substituted or unsubstituted lower alkenyloxy or substituted Or an unsubstituted lower arylaminocarbonylcarbonyl, or R ^5a is taken together to represent an oxygen atom, and X represents an oxygen atom or —CH ₂ —. A certain SH3 domain binding inhibitor according to the above [1] or [ ² ].

 [4] The non-peptidic compound having SH3 domain binding inhibitory activity is represented by the general formula (II):

 (Π)

(In the formula, R ⁶ represents a hydrogen atom, substituted or unsubstituted lower alkyl or substituted or unsubstituted lower alkenyl, and R ⁷ and R ⁹ are the same or different and represent a hydrogen atom, formyl, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower Al Kanoiru or substituted or unsubstituted Ararukiru, and R ¹¹ are the same One or different, a hydrogen atom, a halogen, a hydroxy Doxy, a carboxy, a substituted or unsubstituted lower alkyl, a substituted or unsubstituted lower alkoxy, a substituted or unsubstituted lower alkanol, formyl, cyano, nitro, amino, mono- or di-lower Arukiruamino, a compound represented by the representative) a substituted or unsubstituted lower aralkyl force Noiruamino or substitution or unsubstituted lower alkoxycarbonyl [1] or [2] SH ³ domain binding inhibition described Agent.

 [5] The SH3 domain binding inhibitor according to the above [1] or [2], wherein the non-peptidic compound having SH3 domain binding inhibitory activity is a cytochalasin.

 [6] A non-peptidic compound having SH3 domain binding inhibitory activity is represented by the general formula (Ilia)

 (Ilia)

(Wherein, R ^12a represents a substituted or unsubstituted lower alkyl, a substituted or unsubstituted aryl or a substituted or unsubstituted heteroaryl, Q ¹ represents a single bond or an oxygen atom, and = represents a single bond. Represents a bond or a double bond, and when Q between Q ² and an adjacent carbon atom represents a double bond, Q ² — represents = C (CH ₃ ) —, between Q ² and an adjacent carbon atom. Wherein Q ² represents —C (OH) (CH ₃ ) — when the ^two represent a single bond. The SIB domain binding inhibitor according to the above [1] or [2].

 [7] The non-peptidic compound having SH3 domain binding inhibitory activity is represented by the general formula (Illb)

(Illb) [Wherein, R ^12b has the same meaning as R ^12a , Q ³ and Q ⁵ are the same or different and represent a single bond or an oxygen atom, 2 represents a single bond or a double bond, and 2 Q ⁴ Two represents = C (CH ₃ ), one C (= CH ₂ ) —, one CH (CH ₃ ) — or one C (CH ₃ ) =, and R ^12c and R ^12h are the same or different, Represents a hydrogen atom or hydroxy; R ^12d and R ¹² are the same or different and represent a hydrogen atom or methyl; R ¹² and R ^12g represent formyl; or R ^12f and R ^12g are taken together ---- CR ^12h R ^12g and one CHR ^12e R ^12f are ™ CR ^12h -C (OH) (CH ₃ )-C (= O)-CHR ^12e -or two CR ^12h — A— B — CHR ^12e — (where A and B are the same or different and represent —CH (OH) —, one CH ₂ — or —C (= 0) —). A certain SHB domain binding inhibitor according to the above [1] or [2].

[8] X is an oxygen atom ^,, RR ⁴ and 1 ^{水 素} are hydrogen atoms, R ¹ and R ^3b are the same or different, and are hydroxy, carboxy, substituted or unsubstituted lower alkoxy or substituted or unsubstituted A lower alkanoyloxy, R ^2b is a substituted or unsubstituted lower alkyl, and R ^5a is a group represented by the general formula (IV)

[Wherein R ⁵ represents substituted or unsubstituted lower alkyl, R ^5d and R ⁵ are the same or different and represent a hydrogen atom, hydroxy or substituted or unsubstituted lower alkoxy, or R 5 ^5d and R ^5e represents together a connexion oxygen atom, R ⁵ ¾3 preliminary R represents water atom, formyl, a lower Arukanoiru lower alkyl or substituted or non-replacement of the substituted or unsubstituted, R ^¾ formyl , — CH = NQ (wherein Q is substituted or unsubstituted lower alkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted lower alkylamino, substituted or unsubstituted Arylamino or substituted or unsubstituted arylsulfo Represents nilamino), substituted or unsubstituted lower alkyl or substituted or unsubstituted lower alkyl, and R ^5j is the same or different and represents a hydrogen atom, hydroxy or substituted or unsubstituted lower alkoxy. Or [5] and R ⁵ⁱ together represent an oxygen atom].

 [9] General formula (Va)

 (Va)

(Wherein represents a single bond or a double bond, and R ^12a , Q ¹ and Q ² each have the same meaning as described above) or a pharmacologically acceptable salt thereof.

 [10] General formula (Vb)

 (Vb)

[Wherein, Nini represents a single bond or a double bond, and R ^12b , R ⁱ² \ R ^12d , R ¹² R ^12hヽ Q ⁵ and —— 0-are each as defined above, and Nini CR ^12h — D— CHR ^12e — is two CR ^12h — C (OH) (CH ₃ ) _C (= 0) — CHR ^12e — or CR ^12h —A— B— CHR ^12e — (where A and B are And a pharmacologically acceptable salt thereof.

[11] General formula (VI)

[Wherein, R ^3A and R ^3B are the same or different and each represent a hydrogen atom, hydroxy, hydroxyl, substituted or unsubstituted lower alkoxy or substituted or unsubstituted lower alkanoyloxy, or R ^3A And R ^3B together represent an oxygen atom, represents a substituted or unsubstituted lower alkyl, and R ^5e , R ^5d , R ^5e , R ^5f , RR ⁵ⁱ and R ^5j are as defined above. And R ^5G is formyl, hydroxymethyl, substituted or unsubstituted lower alkoxymethyl, substituted or unsubstituted lower alkyl methoxymethyl, substituted or unsubstituted lower alkyl methoxymethyl, or —CH = NQ ^A

(Wherein Q ^A represents substituted or unsubstituted lower alkoxy, substituted or unsubstituted aryloxy or substituted or unsubstituted aralkyloxy), provided that R ^5G is formyl and R ^3A or R ^{3A When one of 3B} is a hydrogen atom, the other of R ^3A or R ^3B is not hydroxy] or a pharmacologically acceptable salt thereof.

 [12] A medicament comprising the compound of the above-mentioned [9] or [10] or a pharmacologically acceptable salt thereof as an active ingredient.

 [13] A medicament comprising the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof as an active ingredient.

 [14] An SH3 domain binding inhibitor comprising, as an active ingredient, the compound according to the above [9] or [10] or a pharmacologically acceptable salt thereof.

 [15] An SH3 domain binding inhibitor comprising the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof as an active ingredient. -

[16] Any of the above-mentioned [1] to [S], [14] and [I ⁵ ], wherein the SH3 domain binding is a binding between a protein having an SH3 domain and a protein having a proline-rich sequence. 2. The SH3 domain binding inhibitor according to item 1.

 [17] The SH3 domain binding inhibitor according to the above [16], wherein the protein having a SIB. Domain and / or the protein having a proline-rich sequence is a virus-derived protein.

 [18] The SH3 domain binding inhibitor according to the above [17], wherein the virus-derived protein is a protein derived from a retrovirus, a hepatitis virus or a virus.

 [19] Proteins having an SH3 domain are Src, Yes, Fg Hck, Lck ヽ Abl, Fyn, Lyn, Blks Yrk, Ras-GAP, PLC 7 s PI3K :, Tec, Txk / Rlk ヽ Tsk / Emt / Itk, Btk, Crk, Grb2, Neks Vav, STAT, Cortactin, p40-phox, p67-phox, p47-phox ヽ The TCR signal molecule (TCRsm) or the レ 2R? SH3 domain binding inhibitor.

[20] protein with a proline-rich sequence ⁵ , HIV-lNef, p22-phox _N p47-phox, Sam68, Sosl, Dynamin, c-Cbl ヽ Z01, pX ORF I ヽ LHDAg ヽ NS5A ヽ pORF3, ICP10, LMP2A, The $ H3 domain binding inhibitor according to the above [16] or [19], which is Tip or Tio.

 [21] The binding of the protein having the SH3 domain to the protein having the proline rich sequence is Grb2 and Sosl, Fyn and Sam68, Src and Sam68, PLC and Sam68, Grb2 and Sam68, Lyn and HIV-lNef ヽ Hck and HIV -lNef ヽ TCRsm and HIV-lNef ヽ p47-phox and p22-phox ヽ 67-phox and p47-phox, Lyn and Dynamii Cortactin and Z01, Lyn and c-Cbl, IL-2R /? or a-2 and pX ORF I, Grb2 and NS5A ヽ Src and pORF3, Hck and pORF3, Fyn and pORF3, PI3K and p〇RF3, PLC and pORF3, Grb2 and pORF3, Grb2 and ICP10, Lyn and LMP2A ヽ Lck and Tip, Lyn and Tio, The SH3 domain binding inhibitor according to the above [16], which is a bond between Hck and Tio, Lck and Tio, Src and Tio, Fyn and Tio, or Yes and Tio.

[22] The binding between the protein having the SH3 domain and the protein having the proline-rich sequence is Grb2 and Sosl, Fyn and Sam68, Src and Sam68, PLC and Sam68, Grb2 and Sam68, Lyn and HIV-lNef or Cortactin and ZOl. SH3 domain according to the above [16], which is binding Binding inhibitors.

 [23] ^ dUZd ^ used in the production of the compound according to [9] or [10] above, is a filamentous fungus (jylariaks filamentous fungus) MPC1005 (Accession number: FERM BP-7980), Aspergillus species ( A microorganism selected from the group consisting of MPC1006 (Accession number: FERM BP-7899) and Aspergillus sp. (Α "s sp.) MPC1009 (Accession number: FERM BP-7900).

 [24] An antitumor agent comprising the compound of the above-mentioned [9] or [10] or a pharmacologically acceptable salt thereof as an active ingredient.

 [25] An antitumor agent comprising the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof as an active ingredient.

 [26] A therapeutic agent for allergic diseases, comprising as an active ingredient the compound according to the above [9] or [10] or a pharmacologically acceptable salt thereof.

 [27] A therapeutic agent for allergic diseases comprising the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof as an active ingredient.

[28] A therapeutic agent for a viral disease, comprising as an active ingredient the compound according to [ ⁹ ] or [10] or a pharmacologically acceptable salt thereof.

 [29] A therapeutic agent for a viral disease, comprising the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof as an active ingredient.

 [30] An AIDS therapeutic agent comprising the compound of the above-mentioned [9] or [10] or a pharmaceutically acceptable salt thereof as an active ingredient.

 [31] An AIDS therapeutic agent comprising the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof as an active ingredient.

 [32] Use of the compound of the above-mentioned [9] or [10] or a pharmaceutically acceptable salt thereof for the production of an SH3 domain binding inhibitor.

 [33] Use of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof for the production of an SH3 domain binding inhibitor.

[34] The compound according to the above [9] or [10] or a compound thereof for the production of an antitumor agent Use of pharmacologically acceptable salts.

 [35] Use of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof for the manufacture of an antitumor agent.

 [36] Use of the compound of the above-mentioned [9] or [10] or a pharmacologically acceptable salt thereof for the manufacture of a therapeutic agent for an allergic disease.

 [37] Use of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof for the manufacture of a therapeutic agent for an allergic disease.

 [38] Use of the compound of the above-mentioned [9] or [10] or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for a viral disease.

 [39] Use of the compound of the above-mentioned [11] or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for a viral disease.

 [40] Use of the compound or the pharmaceutically acceptable salt thereof according to the above [9] or [10] for the manufacture of a therapeutic agent for AIDS.

 [41] Use of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof for the manufacture of a therapeutic agent for AIDS.

 [42] treatment of various diseases involving SH3 domain binding, characterized by administering an effective amount of the compound according to [9] or [10] or a pharmacologically acceptable salt thereof, or Z or Prevention methods.

 [43] A method for treating and / or preventing or treating various diseases involving SH3 domain binding, which comprises administering an effective amount of the compound according to the above [11] or a pharmacologically acceptable salt thereof.

 [44] A method for treating a malignant tumor, which comprises administering an effective amount of the compound of the above-mentioned [9] or [10] or a pharmacologically acceptable salt thereof.

 [45] A method for treating a malignant tumor, which comprises administering an effective amount of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof.

[46] Treatment and / or prophylaxis of an allergic disease characterized by administering an effective amount of the compound according to [9] or [10] or a pharmacologically acceptable salt thereof. Prevention method.

 [47] A method for treating and preventing or preventing allergic diseases, which comprises administering an effective amount of the compound according to [11] or a pharmacologically acceptable salt thereof.

 [48] A method for treating a viral disease, which comprises administering an effective amount of the compound according to the above [9] or [10] or a physiologically acceptable salt thereof.

 [49] A method for treating a viral disease, which comprises administering an effective amount of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof.

 [50] A method for treating AIDS, which comprises administering an effective amount of the compound of the above-mentioned [9] or [10] or a pharmacologically acceptable salt thereof.

 [51] A method for treating AIDS, which comprises administering an effective amount of the compound of the above-mentioned [11] or a pharmacologically acceptable salt thereof.

 [52] A method for treating AIDS, which comprises administering an effective amount of a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof.

 [53] A method for treating a viral disease, which comprises administering an effective amount of a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof.

 [54] An AIDS therapeutic agent comprising a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof as an active ingredient.

 [55] A therapeutic agent for a viral disease, comprising as an active ingredient a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof.

 [56] Use of a compound having SH3 domain binding inhibitory activity or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for AIDS.

 [57] Use of a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof for the manufacture of a therapeutic agent for a viral disease.

 Hereinafter, the compounds represented by the general formulas (I), (Π), (Ilia), (lift), (Va), (Vb) and (VI) will be referred to as compounds (I), (II), (II) Ilia), (lift), (Va), (Vb) and (VI).

General formulas (I), (Π), (Ilia), (Illb), (IV), (Va), (Vb) and (VI) In the definition of each group of

 (1) Lower alkyl, and lower alkyl of lower alkoxy, lower alkoxycarbonyl, lower alkoxymethyl, lower alkylamino and mono- or di-lower alkylamino include, for example, straight-chain or branched alkyl having 1 to 8 carbon atoms, Specifically, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl and the like, or a cyclic alkyl having 3 to 8 carbon atoms, Specific examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

 (2) The lower alkenyl includes, for example, a straight-chain or branched alkenyl having 2 to 8 carbon atoms, specifically, vinyl, aryl, 1-propenyl, methacrylyl, crotyl,

1-butenyl, 3-butenyl, 2-pentenyl, 4-pentenyl, 2-hexenyl, 5-hexenyl, 1,5-dimethyl-41-hexenyl, etc., or alkenyl having 2 to 4 double bonds, specifically Typically, 1,3-butene genil, 1,3-pentene genil, 2,4-pentene genil, 1,5-dimethyl-1,4-hexagenyl, 1,3,5-hexatrienyl, etc. can give.

 (3) The lower alkanoyl, and the lower alkanoyloxy, lower alkanoyloxymethyl, lower alkanoylmethyl, lower alkanoylamino and lower alkanoylaminocarbonyl portions of the lower alkanoyl moiety include, for example, linear or branched Examples include branched alkanols having 2 to 8 carbon atoms, specifically, acetyl, propionyl, butyryl, isoptyryl, valeryl, isoparryl, piperoyl, hexanoyl, heptanoyl, and oxylnoyl.

 (.4) Examples of aryl, arylaryl, arylsulfonylamino and aryloxy aryl include, for example, aryl having 6 to 14 carbon atoms, specifically, phenyl, naphthyl, anthryl and the like.

(5) The aralkyl and the aralkyl moiety of the aralkyloxy include, for example, aralkyl having 7 to 15 carbon atoms, specifically, benzyl, phenethyl, and benzhydr. Lil, naphthylmethyl and the like.

 (6) The heteroaryl includes, for example, a 5- or 6-membered monocyclic aromatic heterocyclic group containing at least one heteroatom selected from the group consisting of a nitrogen atom, an oxygen atom and a sulfur atom, or a nitrogen atom A bicyclic or tricyclic condensed aromatic heterocyclic group containing at least one heteroatom selected from the group consisting of an oxygen atom and a sulfur atom and having a fused 3- to 8-membered ring. More specifically, for example, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinolinyl, isoquinolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, cinnolinyl, pivalyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, tetrazolyl Oxazolyl, Indolyl, Indazolyl, Ben Imidazoriru, benzotriazoles § benzisoxazolyl, benzothiazolyl, Benzookisazori, purinyl and the like.

 (7) Halogen represents each atom of fluorine, chlorine, bromine and iodine.

 (8) The lower alkenyl part of the lower alkenyloxy has the same meaning as the lower alkenyl (2).

(9) The substituents in the substituted aryl, the substituted aryloxy, the substituted arylamino, the substituted arylsulfonylamino, the substituted aralkyl and the substituted aralkyloxy are the same or different, for example, the number of substitutions:! To 5, hydroxy, halogen, formyl, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkoxy, lower alkanol and the like. The substitution position is not particularly limited. The lower alkyl moiety and the lower alkanoyl of the halogen, lower alkyl and lower alkoxy shown herein are respectively the same as the halogen (7) 7the lower alkyl (1) and the lower alkanoyl (3), and are substituted lower alkyl. The substituent of the lower alkoxy includes, for example, hydroxy or lower alkoxy having 1 to 3 substituents [the lower alkyl portion of the lower alkoxy is the same as the lower alkyl (1)], a halogen [the halogen is the halogen It is the same as (7)]. (10) Substituted lower alkyl, substituted lower alkoxy, substituted lower alkoxycarbonyl, substituted lower alkoxymethyl, substituted lower alkanoyl, substituted lower alkanoyloxy, substituted lower alkanoyloxymethyl, substituted lower alkanoylmethyl, substituted lower alk As the substituents in the nonyloxy, the substituted lower alkanoylamino, the substituted lower alkanoylaminocarbonyloxy, the substituted lower alkylamino and the substituted lower alkenyl, the same or different substituents, e.g., 1 to 3 hydroxy, Lower alkyl, lower alkoxy, carboxy, oxo, amino, epoxy, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylo, etc. Raise It is. The substitution position is not particularly limited. The lower alkanol portion of the lower alkanol and lower alkenyl group, the lower alkyl portion of the lower alkyl and lower alkoxy group, and the aryl portion of the aryl and alkyl groups described herein are respectively the lower alkyl group. It is synonymous with the power nozzle (3), the lower alkyl (1), and the aryl (4). The substituents on the substituted aryl and the substituted aryl have the same meanings as the substituents on the substituted aryl (9), and the substituents on the substituted lower alkanoyl and the substituted lower alkanol include the number of substituents 1 Examples of the above include hydroxy, lower alkoxy [the lower alkyl part of the lower alkoxy is the same as the lower alkyl (1)], halogen [the halogen is the same as the halogen (7)], and the like.

 (11) As cytochalasins, for example, cytochalasin (cytochalasin), oral serichalasin (Rosdlichalasit ^, epoxycytochalasin), force etoglobosin (chaetoglobosin), penocalasin, and aspochalasin. can give.

 (12) SH3 domain binding inhibition refers to inhibition of protein-protein binding via the SH3 domain.

(13) SH3 domain binding may be, for example, any binding between a protein having an SH3 domain and a protein having a proline-rich sequence, for example, SH3 domain. And a protein having a proline rich sequence and / or a protein derived from a virus.

 Examples of proteins derived from the virus include proteins derived from retroviruses such as human immunodeficiency virus-1 (HIV-1) and human T lymphocyte virus-1 (HTLV-1), and Derberian hepatitis virus (HDV). Hepatitis C virus (HCV), Hepatitis E virus

(NEV) and other proteins derived from hepatitis virus, simple herpes virus 1-2 (HSV-2), EB virus (EBV), S. mori herpes virus (H. saimiri), Atererher's virus (H. fltefe) Examples of proteins derived from a herpes virus, such as Src family, include Src, a non-receptor tyrosine kinase Src family protein, and Yes ヽ Fgr ヽ Hck ヽ Lck ヽ Abl ヽ Fyn. Ras-GAP (ras-GTPase-activating) with enzymatic activity such as (fgr / yes-related novel eene Lyn (lgr / yes-related novel gene, Blk (B-cell lymphocyte kinase) _s Yrk, es-related kinase)) protein), PLC

Tec, Txk / Rlk, Tsk / Emt / Itks Btk, a protein of the Tec family of non-receptor-type oral synkinases such as (phospholipase C-gamma) ヽ PI3K (phosphatidylinositol 3-kinase)

STAT, a transcription factor, such as Crk (CT-10bregulated), Grb2 (growth factor receptor -bound protein 2), Nck, and Vav, which are adapter proteins such as (Bruton's tyrosine kinase)

(signal transducers and activators of transcription), etc.Related to the formation of the actino skeleton Cortactin, etc.Signal molecules of T cell receptor, such as p40-phox, p67-phox, p47-phox, etc. TCRsm [Current 'Biology-(Curr. Biol.), Vol. 11, p. 1294 (2001)], etc., the IL-2R (interleukin-2 receptor) chain or chain [Journal of Biology (J. Virol.), Vol. 74, p. 9828

(2000)].

 Proteins having a proline-rich sequence include, for example, p22 which is a component of HIV-lNe, a NefT 'involved in virus infection and / or proliferation, and replication of HIV-1 and a complex of NADPH-oxynase. -Phox, p47-phox, etc.

(src-associated mitotic substrate 68), etc. ZOl related to cytoskeleton, such as Sosl (son of sevenless) which is an adapter protein, Dynamic c-Cbl (casitas B-lineage lymphoma) HTLV-1 pX ORF I (open reading frame I of the pX regione) related to the persistence of virus infection, etc. [Journal of 'ob' Virology 1 (J. Virol.), Vol. 74, p. 9828 (2000) LHDAg (large hepatitis delta antigen) involved in viral RNA synthesis [Journal of Ob 'Virology (J. Virol.), 72, 2089 (1998)], NS5A

[Proceeding. Op. The National Academy of Sciences Op. 'The' U.S.A. (Proc. Natl. Acad. Sci. USA), Vol. 961, 5533 (1999) ], PORF3 [Journal 'Op' Biological 'Chemistry (J. Biol. Chem.), 276, 42389 (2002)] ヽ ICP10 [Journal of Biological Biology Chemistry (J. Biol. Chem.) Biol. Chem.), Vol. 271, p. 17021 (1996)], etc., and LMP2A [Expression of Cell Res.] Related to the latency and Z or permeation of the virus in host cells. 257, ヽ 332 (2000), Journal of Violology (J. Virol.), 69, 7814 (1995)], Tip [Journal of Violology (J. Virol.) ), 72, 2607 (1998):], Tio [Journal-J. Virol., 73, 4631 (1999)], etc. Is raised.

 Examples of the binding between a protein having an SH3 domain and a protein having a proline rich sequence include, for example, Fyn and Sam68 [Journal of Biologic 'Chemistry (J. Biol. Chem.), 272, 6214 (1997). ], Src and Sam68 [Molecular Cell Biology (Mol. Cell Biol.), Vol. 15, 186 (1995)], Grb2 and Sosl [Nature, 363, 83 (1993) )], PLC J and Sam68 [Oncogene, 18 volumes, 4647 pages (1999)], Grb2 and Sam68 [Molecular and Cellular Biology (Mol. Cell. Biol), 15, 186 ( 1995)], Lyn and HIV-lNef or Hck and HIV-lNef [EMBO. J., 14, 484 (1995), Virology, 262, 55 (1999)], TCRsm and HIV-lNef [Curr. Biol., Vol. 11, 1294 (2001)], p47-phox and p22-pho x

[Proceding of the National Academy of Sciences of the _{United States of America} ( _proc . Natl. Acad. Sci. USA), 91, 10650 pages ( 1994)], p67-phox and p47-phox [Proceeding Off, The National Academy Proc. Natl. Acad. Sci. USA, 91, 10650 (1994)], Lyn and Dynamin [Journal 157, 1226 (1996)], Lyn and c-Cbl [Journal 'ob' Biological 'Chemistry (J. Biol. Chem.), 270, 9115 ( 1995)], Cortactin and Zol [Journal of Biological Sciences' Chemistry (J. Biol. Chem.), Vol. 273, p. 29672 (1998)], with the IL-2R /? pX ORF I [Journal of Ob. Virol., Vol. 74, pp. 9828 (2000)], Grb2 and NS5A [Schedule of Op. the National 'Academy' Ob ' Science 'O blanking tHE - Interview one'es' er _{(.... p roc Natl} Acad Sci USA), 961 Certificates, p. 5533 (1999)], Src and pORF3, Hck and pORF3, Fyn and pORF3, PI3K And pORF3, PLCS and pORF3 or Grb2 pORF3 [Journal 'Ob' Biological 'Chemistry (J. Biol. Chem.), vol. 276, p. 42389 (2002)], Grb2 and ICP10 [Journal' Ob 'Biological' Chemistry (J. Biol. Chem.), Volume 271, p. 17021 (1996)], Lyn and LMP2A [Expression Cell Research., Vol. 257, p. 332 (2000), Journal J. Virol., Vol. 69, p. 7814 (1995)], Lck and Tip [Journal op'J. Virol., Vol. 72, p. 2607 (1998) )], Lyn and Tio, Hck and Tio, Lck and Tio, Src and Tio, Fyn and Tio or Yes and Tio [Journal of Virol., 73, 4631 (1999) Year)]. Pharmaceutically acceptable salts of the non-peptide compounds include, for example, pharmacologically acceptable acid addition salts, metal salts, ammonium salts, organic amine addition salts, amino acid addition salts and the like.

Pharmaceutically acceptable acid addition salts of non-peptide compounds include, for example, inorganic acid salts such as hydrochloride, sulfate, nitrate and phosphate, acetate, maleate, fumarate, and citrate Examples of the pharmacologically acceptable metal salts include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, and the like. Aluminum salts, zinc salts, etc. are listed. Pharmacologically acceptable ammonium salts include ammonium and tetramethylammonium. Pharmacologically acceptable organic amine addition salts include morpholine, piperidine and the like; pharmacologically acceptable amino acid addition salts include Addition salts of glycine, phenylalanine, lysine, aspartic acid, glutamic acid and the like can be mentioned.

 Next, a method for producing the non-peptide compound used in the present invention will be described. In the following production methods, if the defined groups change under the reaction conditions or are inappropriate for carrying out the method, methods commonly used in organic synthetic chemistry, such as protection of functional groups, Deprotection, etc. [For example, Protective Groups in Organic Synthesis third edition, TW Greene, John Wiley & Sons, Inc. (John Wiley & Sons Inc.) (1999)]. In addition, the order of the reaction steps such as introduction of a substituent can be changed as necessary.

 The non-peptide compound used in the present invention can be produced, for example, by a series of reactions shown below.

Manufacturing method 1:

 Among the non-peptidic compounds used in the present invention, compound (VI) can be produced by the following series of reaction steps.

Process 1:

Among the compounds (VI), the compound (Via) wherein R ^5G is hydroxymethyl can be produced from the compound (VII) by the following reaction steps.

(式中、 R 、 R^3A、 R^3B、 R^5e、 R^5d、 R^5e、 R^5f、 R、 R⁵ⁱおよび R^5jは、 それぞれ前 記と同義である）

(Wherein, R, R ^3A , R ^3B , R ^5e , R ^5d , R ^5e , R ^5f , R, R ⁵ⁱ and R ^5j have the same ^{meanings as} described above, respectively)

 Compound (Via) can be obtained by treating compound (VII) with a reducing agent in an inert solvent.

 Examples of the reducing agent include sodium borohydride, lithium aluminum hydride, diisobutyl aluminum hydride, sodium bis (2-methoxyethoxy) aluminum hydride, and the like. 1 to: 10 equivalents are used.

 Examples of the inert solvent include methanol, ethanol, chloroform, dichloromethane, tetrahydrofuran (THF), dimethyl ether, toluene, and dimethylformamide (DMF).

 The reaction is carried out at a temperature between 0 ° C. and the boiling point of the solvent used, and is usually completed in 1 minute to 24 hours.

Step 2:

Among the compounds (VI), the compound (VIb) in which R ^5G is a substituted or unsubstituted lower alkoxymethyl or a substituted or unsubstituted lower alkanoyloxymethyl is obtained from the compound (Via) obtained in Step 1 It can be produced by the following reaction steps.

(Wherein, R, R ^3A , R ^3B , R ^5cs R ^5d , R ^5es R ^5f , R, R ⁵ⁱ and R ^{5j each} have the same meaning as described above, and R is a substituted or unsubstituted lower. Alkyl [wherein the lower alkyl has the same meaning as the lower alkyl (1), and the substituent of the substituted lower alkyl has the same meaning as the substituent (10) of the substituted lower alkyl] or a substituted or unsubstituted lower alkyl Canoyl [the lower alkanoyl has the same meaning as the lower alkanoyl (3), and the substituent of the substituted lower alkenyl has the same meaning as the substituent (10) of the substituted lower alkenyl].

The compound (VIb) is the same as the compound (Via) obtained in the step 1:! To 20 equivalents of R—Z (wherein R is as defined above, and Z represents each atom of chlorine, bromine, or iodine) ) In an inert solvent in the presence of a base. When R is a substituted or unsubstituted lower alkanoyl, a carboxylic anhydride in which R ₂ 〇 (where R is as defined above) can be used instead of R—Z.

 Examples of the base include pyridine, triethylamine, potassium carbonate, cesium carbonate, sodium hydride and the like, and the base is used in an amount of 1 to 50 equivalents to compound (Via) or as a solvent.

 Examples of the inert solvent include chloroform, dichloromethane, acetone, dimethyl ether, acetonitrile, and THF ヽ DMF 例 え ば.

 The reaction is carried out at a temperature between 0 ° C. and the boiling point of the solvent used, and is usually completed in 5 minutes to 24 hours.

Step 3:

Of the compound (VI), the compound (Vic) in which R ^5G is —CH = NQ ^A (wherein Q ^A is as defined above) is produced from the compound (VII) by the following reaction step Things can be done.

(Wherein, R, R ^3A , R ^3B , R ^5e , R ^5d , R ^5e , R ^5f , R ^s , R ^5j and Q ^A are as defined above). Compound (Vic) is reacted with compound (VII) by 1 to 10 equivalents of Q ^A NH ₂ (wherein Q ^A is as defined above) in an inert solvent in the presence of a base, if necessary. Can be obtained.

 Examples of the base include pyridine, triethylamine and the like, and the base is used in an amount of 1 to 20 equivalents based on compound (VII).

 Inert solvents include, for example, chloroform, dichloromethane, acetone, methyl ether, acetonitrile, THF, DMF ^.

 The reaction is carried out at a temperature between 0 ° C. and the boiling point of the solvent used, and is usually completed in 5 minutes to 24 hours.

 In addition, the starting material (VII) in this production method 1 is, for example, the Journal of Antibiotics, Vol. 53, p. 579 (2000), or It can be obtained according to the method described in Japanese Unexamined Utility Model Publication No. 58-116686, or according to them. Manufacturing method 2:

 Compound (II) can be produced, for example, from compound (VIII) by the following method.

 (lib)

(Wherein, R ⁶ , R ^w and R ¹¹ have the same meanings as above, respectively, and R and R ⁹ have the same meanings as R ⁷ and above, but do not simultaneously form a hydrogen atom). 1: Compound (Ha) is prepared by reacting compound (VIII) with:! To 10 equivalents of R ⁶ CO ₂ H (wherein is as defined above) or a derivative thereof in an inert solvent in the presence of an acid. Can be obtained.

 Examples of the acid include organic acids such as formic acid, acetic acid, and trifluoroacetic acid; inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid; and Lewis acids such as titanium tetrachloride and boron trifluoride etherate. Among them, boron trifluoride etherate is more preferably used. The acid is used in an amount of 1 to 100 equivalents based on compound (VIII) or as a solvent. Inert solvents include, for example, chloroform, dichloromethane, THF, DMF ^.

 The reaction is carried out at a temperature between -30 ° C and the boiling point of the solvent used, and is usually completed in 1 minute to 24 hours.

 The starting compound (VIII) can be used as a commercially available product or as described in the journal “Ob” American Chemical Chemical Society (J. Am. Chem. Soc.), Vol. 122, p. 3071 (2000). Or it can be obtained according to it.

Step 2:

 Compound (lib) is prepared by treating compound (Ila) obtained in Step 1 of Production Method 2 with 1 to 20 equivalents of RZ (R and Z are as defined above) in an inert solvent in the presence of a base. It can be obtained by reacting.

 Examples of the base include pyridine, triethylamine, potassium carbonate, cesium carbonate, calcium carbonate, sodium hydride and the like, and the base is used in an amount of 1 to 5 equivalents to compound (Ila).

 Examples of the inert solvent include chloroform, dichloromethane, acetone, methyl ether, acetonitrile, and THF ヽ DMF ^.

 The reaction is carried out at a temperature between 0 ° C. and the boiling point of the solvent used, and is usually completed in 5 minutes to 24 hours.

Production method 3:

Compound (Ilia) and compound (Illb) are described in, for example, Japanese Patent Publication No. 10-114776, WO98 / 41205 _s Helvetica 'Himika' Akta (Helv. Chem. Acta), Vol. 62, p. 1501 (1979), etc., or can be obtained according to them.

Production method 4:

 Compound (I) can be prepared, for example, according to the method described in the above-mentioned Production method 1, Journal of America, Chemical Society (Am. Chem. Soc.), Vol. 107, page 256 (1985), Vol. 2549 (1972), etc., The Journal of Antibiotics, 53, 579 (2000), JP-A-58-116686, etc. Method or according to them. Among the non-peptide compounds used in the present invention, compound (Va) and compound (Vb) (cytochalasin derivative) can be produced by the following culture method. Production method 5:

 The cytochalasin derivative is produced by culturing a microorganism having a cytochalasin derivative-producing ability in a medium, producing and accumulating the cytochalasin derivative in the culture, and collecting the cytochalasin derivative from the culture.

 As the microorganism having a cytochalasin derivative-producing ability, any strain can be used as long as it is a strain having a cytochalasin derivative-producing ability. Mutants obtained by mutating these strains by an artificial mutation method, for example, ultraviolet irradiation, 'X-ray irradiation, treatment with a mutagen, etc. Any of these can be used in the present invention.

 Specific examples include AspergiUus sp. MPC1006 and Aspergillus sp. Axperg sp. MPC1009s Black mushroom Xylariales filamentous fungus MPC1005.

In culturing the microorganism having the ability to produce a cytochalasin derivative of the present invention, an ordinary method for culturing a filamentous fungus is applied. As the medium, any of a synthetic medium and a natural medium can be used as long as the medium contains a carbon source, a nitrogen source, an inorganic substance, etc. which can be assimilated by microorganisms. -Carbon sources include glucose, starch, dextrin, mannose, fructo Sucrose, lactose, xylose, arabinose, mannitol, molasses and the like are used alone or in combination. In addition, hydrocarbons, alcohols, organic acids, etc. are used depending on the assimilation ability of the bacteria.

 As a nitrogen source, ammonium chloride, ammonium nitrate, ammonium sulfate, sodium nitrate, urea, peptone, meat extract, yeast extract, dried yeast, corn-steep liquor, soybean powder, casamino acid, etc. are used alone or in combination. Can be In addition, as necessary, sodium chloride, potassium chloride, magnesium sulfate, calcium carbonate, potassium dihydrogen phosphate, magnesium phosphate octahydrate, ferrous sulfate, calcium chloride, manganese sulfate, zinc sulfate, copper sulfate, etc. Inorganic salts can also be added. Furthermore, trace components that promote the growth of the bacteria used and the production of cytochalasin derivatives can be appropriately added.

As a culture method, a liquid culture method, particularly a submerged stirring culture method, is suitable. The cultivation is carried out at a temperature of 16 to 37 ° C, preferably 25 to 32 ° C, at pH 4 to 10, preferably at pH 6 to 8. For adjusting the pH of the medium, aqueous ammonia or ammonium carbonate solution or the like is used. Used. Culture is usually completed in 1 to 10 days, but it is preferable to stop the culture when the cytochalasin derivative is produced and accumulated in the culture solution and in the cells, and the amount of production in the culture reaches the maximum. o Isolation and purification of the cytochalasin derivative accumulated in the culture from the culture are performed according to a method commonly used for isolating and purifying ordinary microbial metabolites from the culture. For example, the culture is separated into a culture filtrate and cells by filtration, and the cell components are extracted from the cells with a solvent such as black form, acetone, and methanol. Then, the extract and the culture filtrate are combined and passed through a polystyrene-based adsorbent, for example, Diaion HP-20 (manufactured by Ryishi Chemical Co., Ltd.), to adsorb the active components, and then, methanol, acetone, or the like is used. Elute. The eluate is concentrated, and a cytolasin derivative is obtained by column chromatography using octyl decyl group-bonded silica gel (ODS), high-performance liquid chromatography, column chromatography using silica gel, or the like. The detection of cytochalasin derivatives during the culturing, isolation, and purification procedures was performed by thin-layer chromatography. It can be carried out by using an Eod reagent.

 The present inventors have proposed that as a strain having cytochalasin derivative-producing ability, MPC1006 and MPC1009, newly isolated from soil and belonging to Aspergillus sp. 'C4¾erg Zi «sp.), Have cytochalasin having SH3 binding inhibitory action. Found to produce derivatives.

 The representative strain MPC1006 producing the compound of the present invention was isolated from soil and has the following bacteriological properties.

 1. Visual observation

 When cultured on malt extract agar at 25 ° C, the diameter of the colonies reaches 38-39 mm on day 7 of culture and 65-68 mm on day 11 of culture. The center of the surface of the colony on day 11 of culture has a very pale reddish yellow color, and the outside has a very pale yellow color. The center of the back of the settlement is light red-yellow at the center and extremely light yellow at the outside.

 When cultured at 25 ° C on potato pour sugar agar, the diameter of the colonies reaches 38 to 40 mm on day 7 of culture and 46 to 48 mm on day 11 of culture. On the 11th day of culture, the center of the surface of the colony is light grayish yellow, and the outside is very pale yellow. It is white in some places. The center of the back of the settlement has a dull yellow color, and the outside has a pale yellow color.

 The growth temperature range of this fungus is 11.5-34 ° C, and it grows best around 28.5 ° C. The growth pH ranges from 3.5 to 11.5 and grows best around pH 7.5.

 2. Light microscopy observation

 The following shows the results of observation of this bacterium with a light microscope when cultured on a malt extract agar medium at 25 ° C for 7 days.

Hyphae have partitions, 1.0-3.0 m wide, smooth, colorless, and well-branched. The conidiophore has no bulkhead, 3.5-5.0 m wide, 420-700 ^ ιη, smooth, colorless and does not branch. The tip of the conidiophore bulges round and forms a cap. Its diameter is 12. ⁵ 25.0 Zm, the entire surface metulae are formed, the length of metulae is 5.5～8.0〃M. At the tip of the metre, a thin, thick-eared phialide (conidia-forming cell) is formed, Is 3.5 to 6.0 m. Before that, conidium is formed from fiaraid into endogenous budding type and fibular type and linked. The filamentous conidium is unicellular, pale yellowish green, spherical to subspherical in shape, and its surface is smooth and 1.5 to 3.0 m in diameter. In this strain, only the anamorph described above is observed, and no teleomorph is observed.

 Further, another representative strain MPC-1009 producing the compound of the present invention was also isolated from the soil, and has the following bacteriological properties.

 1. Visual observation

 When cultured on malt extract agar at 25 ° C, the diameter of colonies reaches 18 to 25 mm on day 4 of culture and 37 to 48 mm on day 11 of culture. On the 11th day of culture, the center of the surface of the colony has a very pale yellow color, and the outside has a yellowish white color. In addition, the back of the settlement is light grayish yellow.

 When cultured at 25 ° C on potato-glucose agar, the diameter of colonies reaches 17-18 mm on the fourth day of culture and 27-31 mm on the 11th day of culture. The center of the colony on the 11th day of culture has a bright gray-yellow surface and a yellowish white outside. The center of the back of the settlement has a light grayish yellow at the center and a very pale yellow outside.

 The growth temperature range of this fungus is 13.0-34.5 ° C, and it grows best around 27.5 ° C. Growth The pH ranges from 3.5 to 11.2 and grows best around pH 7.3.

 2. Light microscopy observation

 The following shows the observation results of this bacterium with an optical microscope when cultured at 25 ° C for 2 weeks on a malt extract agar medium.

Hyphae have septum, width of 1.5-2.5 m, smooth, colorless and well-branched. The conidiophore has no partition, 3.0-5.0 / m wide, 300-350〃m long, smooth, colorless, and does not branch. The tip of the conidiophore bulges round and forms a cap. Its diameter is from 15.0 to 17.5 m, and thin, thick-eared phialides (conidiogenesis cells) are formed on the entire surface. The length of the phiaride is 10.0-11.5 / m. No metres are formed, and the conidia are formed from phialides in an endogenous budding type and a phial type and linked. This Firo-type conidium is unicellular, light green, spherical or subspherical in shape, its surface is smooth, and its diameter is 2.0 to 3.0 m. In this strain, only the anamorph described above is observed, and teleomorph is not observed.

 Based on the above mycological properties, the taxonomic position of the bacterium was determined by The Genera of Fungi Sporulating in Pure Culture, 2nd ed. 2nd ed., Cramer, Vanduz, JA von Arx, 197)), and it was revealed that the thick bacilli eventually belonged to Φ) a filamentous infectious bacterium, Aspergillus sp. Was. The present inventors have named these strains as “Aspergillus sp. MPC1006” and “Aspergillus sp. Deposited as FERM BP-7899 (Original deposit date: February 18, 2002) and FERM BP-7900 (Original deposit date: February 18, 2002) Deposited at Sen-Yu (Tsukuba-Higashi 1-chome 1-1 Central 6th, Ibaraki, Japan).

 In addition, another representative strain MPC1005 producing the compound of the present invention was also isolated from soil, and its bacteriological properties are as follows.

 1. Visual observation

 When cultured at 25 ° C on malt extract agar medium, colonies reach a diameter of 36 to 39 mm on day 7 of culture and 73 to 76 mm on day 11 of culture. On the eleventh day of culture, the center of the surface of the colony is white, and the outside is yellowish white. The center of the back of the settlement is pale yellow, and the outside is very pale yellow.

 When cultured at 25 ° C on potato-budu sugar agar medium, the diameter of colonies reaches 50-55 mm on the 7th day of culture and 75-78 mm on the 11th day of culture. The center of the surface of the colony on day 11 of culture has a yellowish white color, and the outside has a white color. The center of the back of the settlement has a dull reddish yellow color, and the outside has a very pale yellow color.

 The growth temperature range of this fungus is 13.3-38.6 ° C, and it grows best around 33.0 ° C. Growth The pH ranges from 3.7 to 9.4 and grows best around pH 7.2.

2. Light microscopy observation The following shows the results of observation of this bacterium with a light microscope when cultured on a malt extract agar medium at 25 ° C for 4 days.

 Hyphae have septum, width 0.5-3.0 m, smooth, colorless, sometimes colored and well-branched.

 Using both agar media described above, the cells were cultured at 25 ° C for 2 months or more. In this generation, neither anamorph nor teleomorph was observed.

3. Other 'I' substrate

 The partial nucleotide sequence (1228 bp) of the 18S ribosomal DNA (18S rDNA) of this bacterium had the nucleotide sequence represented by SEQ ID NO: 1.

 Using the above base sequence and the 18SrRNA or 18SrDNA sequence of a known fungus, molecular phylogenetic analysis was performed by the neighbor-joining method [Program name: CLUSTAI W, Bulletin of the Japanese Society for Microbial Ecology, Vol. 10, p. 119 (1995)]. As a result, the bacterium belonged to the fungus Pyronomycetes and was closely related to the cyst fungus (Xylariales).

 Based on the above results, the present inventors named this bacterium “Black-spotted mushroom, Nippon-yaki orchid MPC1005 (Xylariales filamentous fungus MPC1005)”, and received the accession number FERM BP-7980 (Original deposit date: March 2002) 26th) Deposited at the National Institute of Advanced Industrial Science and Technology (AIST) Patent Depositary Center (1-1 Tsukuba East, Ibaraki Prefecture, Japan, 1 Chuo No. 6)

7 o

Furthermore, conversion of each functional group in compounds (I), (II), (Ilia), (Illb), (Va), (Vb), (VI) and the starting compound, and conversion of functional groups contained in the substituents In addition to the above-mentioned steps, other methods known in general in synthetic organic chemistry, such as oxidation, reduction, hydrolysis, and other known methods [for example, Comprehensive Organic Transformation 3rd Edition (Comprehensive Organic Transformations 3rd Edition) RC Larock, John Wiley & Sons Inc. (1999), etc.], and the like. However, the order of the reaction steps can be changed if necessary. The intermediates and target compounds in each of the above production methods are isolated and purified by separation and purification methods commonly used in organic synthetic chemistry, for example, filtration, extraction, washing, drying, concentration, recrystallization, various chromatographies, etc. can do. In addition, the intermediate can be subjected to the next reaction without particular purification.

 Compound (I), (II), (Ilia), (Illb), (Va), (Vb) or (VI) may have stereoisomers such as geometric isomers and optical isomers However, all the possible isomers and mixtures thereof, including these, are also included in these compounds, and the non-peptidic conjugates used in the present invention, compounds (1), (11), (Ilia ), (IIIb), (Va), (Vb) or (VI), the non-peptidic compound used in the present invention, compound (I), (本), (Ilia), When (Illb), (Va), (Vb) or (VI) is obtained in the form of a salt, it may be purified as it is, and when it is obtained in the free form, the non-peptide compound used in the present invention may be used. Compound, compound (1), (11), (Ilia),

(Illb), (Va), (Vb) or (VI) may be dissolved or suspended in an appropriate solvent, and then isolated or purified by adding an acid or a base.

 In addition, the non-peptide compound used in the present invention, compound (I), (II), (Ilia), (Illb), (Va), (Vb) or (VI) and its pharmacologically acceptable The salt may be present in the form of an adduct with water or various solvents, and these adducts are also included in the above-mentioned conjugate or a pharmaceutically acceptable salt thereof.

Specific examples of the compound obtained by the present invention are shown in Tables 1 to 3 as compound (Va), (Vb) or (VI).

Table 1

 Table 2 Compound (Va)

第 3表 化合物 (Vb) Compound 16 Compound 17

Table 3 Compounds (Vb)

本発明では上記第 1〜 3表にあげた化合物も使用されるが、 それらの他に本発 明で使用される化合物の具体例を、 化合物 (I) 、 (Π) 、 (Ilia) または （Illb) として第 4〜 7表に示す。

In the present invention, the compounds shown in the above Tables 1 to 3 are also used, and specific examples of the compounds used in the present invention include compounds (I), (Π), (Ilia) and (Ilia). Illb) is shown in Tables 4-7.

Compound R ¹ , R ^3a R ^5f , R ^5h R ^5g

1

 -OH H = 0 -CHO

(UCS15A)

5-OH H = 0-GH = NNHGH ₃ Table 5

Compound R ^12a

12 (Me `` WF1726) -OCH ₃

13 (Cytochalasin E) H

Table 7 Compounds (Illb)

 Next, the pharmacological action of the representative compounds (I), (II), (Ilia), (Illb), (Va), (Vb) or (VI) will be specifically described with reference to test examples.

Test example 1: SH3 domain binding inhibition test 1

(1) Reaction of a protein containing a proline-rich sequence with a compound Human recombinant Sam68 (hereinafter referred to as Sam68AC) containing two proline-rich sequences at the C-terminal side of the five proline-rich sequences in the protein Sam68 having a proline-rich sequence [Sam68 (331-433); RSB (10 mmol / L Tris monohydrochloride, pH7) containing 0.25 g of 0.1% BSA (bovine serum albumiii) [bovine blood albumin (FV); Nacalai Tesque, Inc.] .6, 10 mmol / L NaCl, 1.5 mmol / L MgCl ₂ ) In addition to ImL, a Sam68 ΔC solution was prepared. The compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mmol / L DMSO solution. The DMSO solution was added to the above Sam68ΔC solution such that the final concentrations were 20 and 100 Hmol / L, respectively. The amount of DMSO added was the same. The Sam68A C solution containing the compound was incubated at 37 ° C for 6 hours while gently rotating in a rotary incubator to remove the compound.

Reacted with Sam68 △ C.

 (2) Reaction of protein containing SH3 domain with protein containing proline-rich sequence after compound addition

Fyn-SH3 protein-agarose beads (hereinafter referred to as Fyn-SH3 beads) in which the mouse recombinant SH3 domain of protein SH3 having SH3 domain is bound to agarose beads are added to the Sam68AC solution reacted with the compound in (1) above. 85-139) AC; manufactured by Santa Crutz ']. The protein-protein binding reaction between Sam68ΔC after the compound reaction and Fyn-SH3 beads containing the SH3 domain was carried out at 4 ° C for 16 hours while gently rotating in a rotary culture machine. After the reaction, the complex of Sam68 △ C and Fyn-SH3 beads was precipitated by a rotary centrifuge at 4 ° C and 50 ppm for 3 minutes, and then a TritonX / Np40 extraction buffer solution containing a surfactant [50 mmol / L Tris-1 Hydrochloric acid pH 7.4, 150 mmol / L NaCl lmmol / L ethylenediaminetetraacetate (EDTA), 1% Triton X-100 (TritonX-100), 0.5% nonidet P-40 (NP-40)] Add ImL and add 4 ° Rotation at C for 10 minutes After gentle rotation in the incubator, the same precipitation and washing was performed twice. To the complex of Sam68AC and Fyn-SH3 beads after washing, add ³⁰ L of Laemmli sample buffer [see Chemical Chemistry of Protein I-Separation and Purification, Tokyo Chemical Dojin, p. 211 (1976)]. After mixing well, the mixture was heated at 100 ° C for 10 minutes. After precipitating the beads at 100 ppm in a rotary centrifuge at 25 ° C for 5 minutes, the supernatant was subjected to 10% polyacrylamide gel electrophoresis.

(3) Detection of Sam68AC bound to Fyn-SH3 by Western plot Sam68A C in the supernatant was detected by Western blot as follows. First, the gel after electrophoresis in the above (2) was plotted on a nitrocellulose membrane (Protran; Schleicher and Schuel 0) having a pore size of 0.45 μm. Place on phosphate buffer (PBS) containing 0.25% gelatin and 0.2% Tween-20 (hereinafter, referred to as PBS-TG) and place at 4 ° C overnight to block non-specific binding. Hose-radish peroxidase (HRP) -conjugated mouse anti-GST antibody [GST (B-14) HRP; manufactured by Santa Crutz] diluted 1: 1000 with TG was reacted for 2 hours. after washing with PBS (hereinafter referred to as PBS-T) containing 0.2% T _W een-20, detection was carried out by chemiluminescence had use ECL reagent (Amersham Pharmacia 'Biotech).

 As a result, as shown in FIGS. 1 to 3, Sam68A C bound to Fyn-SH3 beads in the supernatant in a concentration-dependent manner by, for example, compound (I), (11), (Ilia) or (VI). It has been confirmed that the amount of phenol has decreased.

Test Example 2: SH3 domain binding inhibition test 2

 (1) Preparation of cell extract

え、 よく混合させた後、 回転培養機 でゆるやかに回転させながら 4°Cで 45分間攪拌して、 細胞質内蛋白質を抽出した。 この細胞抽出液を 4°C、 15,000rpmで 30分間遠心し、 上清を回収した。 Human colon cancer-derived cell line HCT116 (ATCC No. CCL-247) was used at 37 ° C. using a modified McCoy's medium [McCoy's 5A modified medium; Gibco (GIBCO)] containing 10% ゥ, fetal serum. in 5% C0 ₂ under the conditions of CC T Nkyube Isseki one were cultured in 100mm diameter Didzushi Interview for cell culture. Then, compound 1 was added to two dishes of HCT116 cells on the second day so that the final concentrations were 0.5, 1, 2, 2.5, 5, 10, 20, and 30 mol / L, respectively. . Compound 1 to be added was previously diluted with DMSO so that the amount of DMSO added was the same. HCT116 cell tissue with Compound 1 was added at 37 ° C! , And incubated 5% C0 ₂ under the conditions of the C0 ₂ incubator base Isseki one 2 hours, allowed to act compound. Collect two dishes of HCT116 cells with compound 1 and react with the medium.

After mixing well, the mixture was stirred at 4 ° C for 45 minutes while gently rotating in a rotary culture machine to extract cytoplasmic proteins. The cell extract was centrifuged at 15,000 rpm at 4 ° C for 30 minutes, and the supernatant was collected.

 (2) Immunoprecipitation of SH3 domain-binding protein complex with antibodies

 The cell extract obtained after the addition of the compound obtained in the above (1) was added with 3 g of anti-Src antibody [manufactured by Santa Crutz] and anti-PLC antibody [Santa Crutz] 3〃g, anti-Grb2 antibody [from Sanyo Cruz (5 && 1 ^)] 3 ^, or anti-Cortactin antibody [from Sanyo Cruz (Santa Cmtz)] 3〃g The reaction was carried out at 4 ° C for 16 hours with gentle rotation. Protein A / G-conjugated agarose beads 30 were added to each SH3 domain-binding protein complex after the antibody reaction, and reacted at 4 ° C for 1 hour while gently rotating in a rotary culture machine. After the reaction, precipitate the SH3 domain-binding protein complex with a rotary centrifuge at 4 ° C, 50 ppm for 3 minutes, add lmL of TritonX / Np40 extraction buffer containing detergent, and spin culture at 4 ° C for 10 minutes. After gentle rotation on the machine, precipitation and washing were performed three times in the same manner. After washing, add the SH3 domain-binding protein bead complex and 30 µL of Laemmli sample buffer [Protein Chemistry I-Separation and Purification, Tokyo Kagaku Dojin, p. 211 (1976)], and mix well. Heated at 10 CTC for 10 minutes. After precipitating the beads for 5 minutes at 25 ° OOOOppm in a rotary centrifuge, the supernatants were each subjected to 7.5% polyacrylamide gel electrophoresis.

 (3) Detection of protein containing prolinrich sequence bound to protein containing SH3 domain by Western plot

A protein containing a proline-rich sequence in each SH3 domain-binding protein complex in the supernatant was detected by Western blotting as follows. First, the protein in the gel after the electrophoresis described in (2) above was plotted on a 0.45 µm nitrocellulose membrane [Protran; Produced by Schleicher and SchuelO], and PBS was applied to the membrane. Non-specific binding was blocked by loading -TG. Eg anti-Sam68 antibody for detection of Sam68, Eg anti-Sosl antibody for detection of Sos, and detection of Z01 for Z01. ゥ A heron anti-ZOl antibody was diluted 1: 1000 with PBS-TG and reacted for 2 hours. After washing with PBS-T, an HRP-conjugated anti-Egret antibody (Amersham) diluted 1: 4000 with PBS-TG was reacted for 1 hour. After washing with PBS-T, detection was performed by chemiluminescence using an ECL reagent (Amersham Pharmacia Biotech). As a result, as shown in FIGS. 4 to 7, as shown in FIG. 4, the amount of Sam68 bound to the Src, and the amount of Sam68 bound to the PLC in FIG. In Figure 6, the amount of Sam68 bound to Grb2, the amount of Sosl bound to Grb2 in Figure 6, and the amount of Z01 bound to Cortactin in Figure こ と が were confirmed to be reduced in a concentration-dependent manner by the addition of compound. Was.

Test Example 3: SH3 domain binding inhibition test 3

 (1) Reaction of protein Nyn containing proline-rich sequence and protein Lyn with SH3 domain with compounds

Recombinant Nef derived from HIV-1 having a proline-rich sequence [Nef (3-190); manufactured by Cortex Corporation] 0.3 μg and purified Lyn derived from sea bream [Lyn; manufactured by Upstate Biotechnology] 1 Zg was added to 1 mL of RSB (10 mmol / L Tris-HCl pH7.6, 10 mmol / L NaCi .5 mmol / L MgCl ₂ ), a hypotonic buffer containing 0.1% BSA (bovine serum albumin), and Nef And a mixed solution of Lyn was prepared. The compound was dissolved in DMSO to prepare a DMSO solution having a concentration of 10 mmol / L. The DMSO solution was added to the above-mentioned mixed solution of Nef and Lyn such that the final concentrations were 1, 2, 5, and 10 mol / L, respectively. The compound to be added was diluted in advance with DMSO so that the amount of DMSO added was the same. The mixed solution of Nef and Lyn to which the compound had been added was stirred at 4 ° C for 16 hours while gently rotating in a rotary culture machine to react the compound with Nef and Lyn proteins.

 (2) Immunoprecipitation of SH3 domain-binding protein complex with antibodies

To the mixed solution of Nef and Lyn obtained after adding the compound obtained in the above (1), add l〃g of anti-Lynibody [manufactured by Santa Cmtz] and gently rotate in a rotary culture machine. The reaction was continued at 4 ° C for 16 hours. SH3 domain binding protein of Nef / Lvn after antibody reaction 10 μL of protein A / G-bound agarose beads was added to the complex, and the mixture was reacted at 4 ° C. for 1 hour while gently rotating in a rotary culture machine. After the reaction, precipitate the SH3 domain-binding protein complex with a rotary centrifuge at 35 ° C for 3 minutes at 4 ° C, add ImL of TritonX / NP40 extraction buffer containing detergent, and spin culture at 4 ° C for 10 minutes. After gentle rotation on the machine, precipitation and washing were performed three times in the same manner. To the Nef_Lyn SH3 domain-binding protein bead complex after washing, add 30 L of Laemmli's sample buffer [Evening protein chemistry I-Separation and purification, Tokyo Chemical Dojin, p. 211 (1976)] After mixing well, the mixture was heated at 100 ° C for 5 minutes. After precipitating the beads at 25 ° C. in a rotary centrifuge at 100 rpm for 5 minutes, the supernatant was subjected to 7% polyacrylamide gel electrophoresis.

 (3) Detection of Nef protein containing prolinrich sequence bound to Lyn protein containing SH3 domain by Western protocol

 The Nef protein containing a proline-rich sequence in the Nef / Lyn SH3 domain-binding protein complex in the above supernatant was detected by Western blotting as follows. First, the protein in the gel after the electrophoresis in the above (2) was plated on a 0.45 µm nitrocellulose membrane [Protran; Produced by Schleicher aixi Schuell], and then applied to the membrane. Non-specific binding was blocked with PBS-TG. Since galactosidase was fused to the recombinant Nef protein used, Nef was detected using an anti-galactosidase antibody (anti-^-gal antibody). That is, a heron anti-galactosidase antibody was diluted 1: 1000 with PBS-TG and reacted for 2 hours. After washing with PBS-T, an HRP-conjugated anti-Egret antibody [manufactured by Amersham] diluted 1: 4000 with PBS-TG was reacted for 1 hour. After washing with PBS-T, detection was performed by chemiluminescence using an ECL reagent (Amersham Pharmacia Biotech).

 As a result, as shown in FIG. 8, it was confirmed that the amount of the Nef protein containing the proline-rich sequence bound to the Lyn protein containing the SH3 domain was reduced in a concentration-dependent manner by the addition of the compound. .

Test example: Calculation of the SH3 domain binding inhibition rate of a compound Based on the results plotted according to the method of Test Example 1, the SH3 domain binding inhibition rate of the compound was calculated as follows. The band intensity of Fyn-1 SH3 and Sam ⁶⁸ AC developed on the film by chemiluminescence was measured using a gel scanner-1 (manufactured by TOYOBO). Next, the band intensity of Sam68AC was divided by the band intensity of Fyn-SH3 to calculate the band intensity of Sam68AC with respect to the band intensity of Fyn-SH3. The ratio of the band intensity of Sam68AC to the band intensity of Fyn-SH3 when no compound was added was defined as 100%, and the ratio of decrease in the band intensity of Sam68AC after the compound was added was defined as the inhibition ratio of SH3 domain binding of the compound. The results are shown in Table 8.

Compound SH3 domain binding inhibition rate (%) Test compound concentration (μmol / L)

Compound

 0 20 60 100 150

1 0 0 46

2 0 34 95

 3 0 75 100

 4 0 47 85

 5 0 77 100

 6 0 28 37

 H 0 82

8 0 77

9 0 90

10 0 71

1 1 0 84

12 0 39 88

 13 0 0 21

 14 0 80

 15 0 39

 16 0 67 78

17 0 36

18 0 16 67

19 0 40

20 0 92 100

21 0 64 100

22 0 91 100

23 0 82

24 0 62 80

25 0 87 100

26 0 92 100

27 0 100 100

28 0 31 73 As described above, according to Test Examples 1 to 4, Compounds (I), (II), (Ilia), (Illb), (Va), (Vb) and (VI) exhibited excellent SH3 domain binding inhibitory effects.

Test Example 5: HIV-1 growth suppression test

 (1) HIV-1 infection and growth suppression experiment

MT2 cells (T cell line) were transformed using RPMI1640 medium [Gibco (GIBCO) No. 26140-076] containing 10% fetal bovine serum [GIBCO (No. ²⁶ 140-079)]. The volume was adjusted to 10 ⁵ cells / mL, and 1 mL was dispensed per well into a 24-well plate [24 well plate: No.3526, manufactured by Coster Corporation]. The HIV-1 LAI strain {DNA clone pLAI including the entire genome [GenBank accession No. K02013, Virology, 185, 661 (1991)] was obtained from FuGENE6 Transfection Reagent (Roche). , No. 1814443], transfected into Hela ffl cells, and inoculated with the culture supernatant from day 2 to a final concentration of 500 cpm / iL in each of the above cells. The sample was allowed to stand still at ° C (virus infection). Here, compound or PP2 (produced by Alexs¾, manufactured by Protin Phospho-Nose 2: 4-amino-5- (4-chlorophenyl) -7- (tert-butyl) pyrazo [3,4-d] pyrimidine, Src Inhibitors of the phosphorylation of lip synthase (control) were added to give final concentrations of 1, 5, 10, 25 and 50 mol / L (drug addition). Thereafter, after infection with the virus, culture supernatants of 50 / L were collected at 3, 5, 7, and 9, respectively, and stored at -80 ° C until measurement. At this time, the virus which infected the virus and did not add the drug was defined as positive control, and the virus which did not infect the virus and did not add the drug was defined as negative control. The test was performed three times with the same contents.

 (2) Measurement of HIV-1 level

 The quantification of HIV-1 was determined by the RT assay method (Reverse) with a slight modification to the method described in Journal Off, J. Virol., Vol. 38, p. 239 (1981). Transcriptase assay).

To 5 µL of the culture supernatant obtained in the above (1), add RT cocktail (50 mmol / L Tris pH 7.5, 50 mmol / L EDTA ヽ 75 mmol / L KC1, 5 mmol / L MgC, 5 jLL g / mL Poly (A ) [Pharmacia (Pharmacia) No.27-4110-01], 1.6〃g / mL oligo (dT) 12-18 [Pharmacia No.27-7858-01] ヽ 5mmol / L DTT (dithiolate), 0.05% NP-40, 10 / Ci / mL [ shed - ³² P] dTTP} was added 25〃L and incubated for 2 hours at 37 ° C. After the reaction, the reaction solution 6〃L of their DEAE filtermat [Perkin Elmer one (PerMn Elmer) Co., Nanba1450-522] spotted, after removing the shed one ³² P] dTTP unreacted Infiltrate MeltiLex-A [Perkin Elmer, No. 1450-441], and measure the amount of HIV-1 as RT activity count (cpm / L) in MICROBETA The measurement was performed using a PLUS liquid scintillation counter [Perkin Elmer Co.].

 As a result, as shown in Figs. 12 to 14, in the system to which PP2 was added as a control, the amount of HIV-1 increased as in the case of the positive control, and as shown in Figs. It was confirmed that in the system to which the compound was added, the increase in the amount of HIV-1 was suppressed in a concentration-dependent manner. In other words, the effect of suppressing the proliferation of HIV-1 was observed by adding the compound.

Test Example 6: Cytotoxicity test

MT2 cells (T cell line) were transformed into 2x using RPMI1640 medium [manufactured by Gibco (GIBCO), No. 26140-076] containing 10% fetal bovine serum [GIBCO (No. 26140-079)]. It was adjusted to 10 ⁵ cells / mL, 96 Werupureto [96 well plate: Costar (Coster) Co.] was dispensed at 100 L per minute Kakuu Ell. After adding the compound to each cell and culturing, observe the state of the cells on the third day, collect the cells, stain the cells with 0.4% trypan blue [SIGMA], Τ-8154]. The number of surviving cells was counted. As a result, even when the compound 1 was added at a concentration of 10 mol / L, no decrease in the number of viable cells was observed.

As described above, according to Test Examples 1 to 6, it was found that a drug containing a compound having an excellent SH3 domain binding inhibitory activity as an active ingredient can be used for various diseases involving protein-protein binding via SH3 domain (e.g., AIDS, malignant tumor, allergy) It is suggested to be effective for sexual diseases, viral diseases, etc. [Biopolymer, vol. 43, p. 383 (1997)]. Compound (I), (II), (Ilia), (Illb), (Va), (Vb) or (VI) or a pharmacologically acceptable salt thereof can be administered alone as it is. However, it is usually desirable to provide them as various pharmaceutical preparations. The pharmaceutical preparations are used for animals and humans.

 The pharmaceutical preparation according to the present invention comprises a compound (1), (11), (Ilia), (IIIb), (Va), (Vb) or (VI) as an active ingredient or a pharmacologically acceptable salt thereof. Can be contained alone or as a mixture with any other therapeutically active ingredient. In addition, such pharmaceutical preparations are produced by mixing the active ingredient with one or more pharmacologically acceptable carriers and by any method well known in the technical field of pharmaceuticals.

 It is desirable to use the most effective route for the treatment, for example, oral or parenteral such as intravenous.

 Examples of the administration form include tablets, powders, granules, syrups, and injections. Liquid preparations suitable for oral administration, such as syrups, include, for example, water, sugars such as sucrose, sorbitol, fructose, glycols such as polyethylene glycol and propylene glycol, oils such as sesame oil, olive oil, and soybean oil. Preservatives such as p-hydroxybenzoic acid esters, flavors such as strawberry flavor and peppermint

—Can be manufactured using such as Tablets, powders, granules and the like include excipients such as lactose, pudose, sucrose and mannitol, starch, disintegrants such as sodium alginate, lubricants such as magnesium stearate, talc, poly It can be produced using a binder such as vinyl alcohol, hydroxypropylcellulose, and gelatin, a surfactant such as a fatty acid ester, and a plasticizer such as glycerin.

 Formulations suitable for parenteral administration comprise a sterile aqueous preparation containing the active compound which is preferably isotonic with the blood of the recipient. For example, in the case of an injection, a solution for injection is prepared using, for example, a carrier comprising a salt solution, a glucose solution or a mixture of a salt solution and a butudose solution.

Also, in these parenteral preparations, the diluents, preservatives, and flavoring agents exemplified for the oral preparations are used. One or more auxiliary ingredients selected from bars, excipients, disintegrants, lubricants, binders, surfactants, plasticizers and the like can also be added.

The dose and frequency of compound (I), (II), (Ilia), (Illb), (Va), (Vb) or (VI), or a pharmaceutically acceptable salt thereof, It depends on the form of administration, the age and weight of the patient, the nature or severity of the condition to be treated, etc., but in the case of oral administration, usually 0.01 mg to lg per adult, preferably 0.05 to 50 mg once or more per day Dosing once. In the case of parenteral administration such as intravenous administration, 0.001 to 100 _mg , preferably 0.01 to 10 _mg , per adult is administered once to several times a day. However, the dose and the number of administrations vary depending on the various conditions described above.

i i simple simple PHP 昍

 FIG. 1 shows the results of an assay for inhibiting the binding of Sam68 C and Fyn-SH3 at the in-vitro mouth when compounds (I) and (VI) were added at different concentrations. The numbers at the top of each lane indicate the compound number and the concentration (Lmol / L) of each compound added to the Sam68AC solution. The description on the left shows the name of each plot.

 FIG. 2 shows the results of inhibition of binding between Sam68AC and Fyn-SH3 at the in-vitro mouth when compound 1 and compound (Ilia) were added at different concentrations. The numbers at the top of each lane indicate the compound number and the concentration (〃mol / L) of each compound added to the Sam68 ΔC solution. The description on the left shows the name of each plot.

 FIG. 3 shows the in vitro results of the inhibition of the binding of Sam68AC and Fyn-SH3 when the compounds (II) and (Ilia) were added at different concentrations. The numbers above each lane indicate the compound number and the concentration (^ mol / L) of each compound added to the Sam68AC solution. The description on the left shows the name of each plot.

FIG. 4 shows the results of a binding inhibition assay between Src and Sam68 in cells when Compound 1 was added at different concentrations. The numbers at the top of each lane indicate the concentration (/ mol / L) of compound 1 added to HCT116 cells. The description on the left shows the name of the broth. FIG. 5 shows the results of the binding inhibition assay between the intracellular PLC and Sam68 when compound 1 was added at different concentrations. The numbers at the top of each lane are for HCT116 cells. Shows the concentration (md / L) of Compound 1 added. The description on the left shows the name of the plot. FIG. 6 shows the results of assays for inhibiting the binding of Grb2 to Sosl in cells when compound 1 was added at different concentrations. The numbers at the top of each lane indicate the concentration (Zmol / L) of compound 1 added to HCT116 cells. The description on the left shows the name of the broth. FIG. 7 shows the results of the assay for inhibiting the binding of intracellular Cortactin and ZOl when the compound 1 was added at a different concentration. The numbers above each lane indicate the concentration (mol / L) of compound 1 added to HCT116 cells. The description on the left shows the name of the plot. FIG. 8 shows the results of in vitro binding inhibition assay of Nef and Lyn when compounds 1 and 3 were added at different concentrations. The numbers at the top of each lane indicate the compound number and the concentration (〃mol / L) of the compound added to the mixed solution of Nef and Lyn. The description on the left shows the name of the plot.

 Figures 9 to L1 show the results of the HIV-1 proliferation inhibition test (RT Assay) when compound 1 was added at different concentrations. The results were obtained by performing three tests under the same conditions. is there. The left side of the vertical axis of the graph shows the amount of HIV-1 as the count of RT activity (cpmA L), and the lower side of the horizontal axis of the graph shows the number of days elapsed since HIV-1 infection (days). Is shown. The meaning of each plot on the graph is as follows.

U: Positive contorol, X: Negative contorol, ·: Diagonal compound 1 (50 Zmol / w, ▲: Compound 1 (25 Zmol / L), Country: Compound 1 (10 mol / L), 〇: Compound 1 ( 5〃moI / L), △: Compound 1 (l / mol / L)

 Figs. 12 to 14 show the results of the HIV-1 growth inhibition test (RT Assay) when PP2 was added at different concentrations, and the results of three tests performed under the same conditions. is there. On the left side of the graph, the amount of HIV-1 is shown as RT activity count (cpm / 〃L). On the lower side of the graph, the number of days elapsed since HIV-1 infection was plotted. (Day). The meaning of each plot on the graph is as follows.

U: positive contorol, X: negative contorol, arrival: PP2 (, 50 Zmol / L),

 ▲: PP2 (25 mol / L), ■: PP2 (10 mol / L), 〇: PP2 (5 Zmol / L),

△: PP2 (l ^ mol / L) Day S »own

 Hereinafter, embodiments of the present invention will be described with reference to Examples and Reference Examples.

Example 1: Tablet

A tablet having the following composition is prepared by a conventional method. Compound 2 (40 g), lactose (286.8 g) and potato starch (60 g) are mixed, and a 10% aqueous solution of hydroxypropyl cellulose (120 g) is added thereto. This mixture is kneaded by a conventional method, granulated, dried, and then sized to obtain granules for tableting. To this was added and mixed Maguneshiumu stearate (1.2 g), a tableting machine having a pestle of diameter 8mm in performing tableting (Kikusui, RT-15), tablets (per tablet active ingredient ² Omg Containing).

Formulation Compound 2 20 mg

 Lactose 143.4 mg

 30 mg

 Hydroxypropyl cellulose 6 mg

 0.6 mg of magnesium diphosphate

 200 mg

Example 2: Capsule

 A capsule having the following composition is prepared by a conventional method. Compound 12 (200 g), Avicel (995 g) and magnesium stearate (5 g) are mixed by a conventional method. This mixture is filled into Hard Capsule No. 4 (capacity of 120 mg per capsule) with a capsule filling machine (LZ-64 type, manufactured by Zanasi) to obtain a capsule (containing 20 mg of active ingredient per capsule). .

Formulation 1 2 20 mg

 Avicel 99.5 mg

 Magnesium stearate 0.5 mg

 120 mg

Example 3: Injection

An injection having the following composition is prepared by a conventional method. Purify Compound 10 (lg) Dissolve in soybean oil and add purified egg yolk lecithin (12 g) and glycerin for injection (25 g). This mixture is kneaded and emulsified with distilled water for injection to 100 mL in a conventional manner. The resulting dispersion is aseptically filtered using a 0.2 / m disposable membrane filter, and then aseptically filled into glass vials in 2 mL increments. Injection (containing 2 mg of active ingredient per vial) Get.

Formulation Compound 102 mg

 Refined soybean oil 200 mg

 24 mg

 Glycerin for injection 50 mg

 For distillation k 1.72 mL

 2.00 mL

Example 4: Compound 2

 Compound 1 (20 mg; 0.042 mmol) obtained in Reference Example 1 was dissolved in THF (2 mL), and a 70% sodium bis (2-methoxyethoxy) aluminum hydride in toluene (0.040 mL) solution was cooled with ice. added. After stirring at 0 ° C for 20 minutes, lmol / L acetic acid-THF solution (ImL) and ethyl acetate (10mL) were sequentially added to the reaction solution for extraction, and the organic layer was washed with water and lmol / g acid water. . The organic layer was dried over anhydrous sodium sulfate and concentrated, and the obtained residue was crystallized from a mixed solvent of diisopropyl ether (IPE) and n-hexane to obtain 12.3 mg of Compound 2 (yield 61%).

Compound 2:

¾-NMR (CDC1 ₃ ) δ (ppm): 0.78 (3H, d, J = 6.6 Hz), 0.86 (3H, t, J = 7.3 Hz), 1.02 (3H, d, J = 6.6 Hz), 1.07 ( 3H, t, J = 7.7Hz), 1.23 (2H, m), 1.52-1.61 (3H, m), 1.79 (2H, m), 1.96-2.01 (2H, m), 2.66 (IH, d, J = 2.9Hz), 3.27 (IH, t, J = 6.1Hz), 3.39 (3H, s), 3.54 (1H, m), 3.92 (1H, d, J = 7.7Hz), 4.16 (1H, m), 4.35 (1H, brt, J = 9.9Hz), 4.87 (2H, s), 4.97 (IH, d, J = 2.8Hz), 7.41 (IH, s), 9.24 (IH, s), 13.4 (IH, s) ; FAB-MS m / z 505 [M + Na] ⁺ .

Example 5: Compound 3 Compound 2 (10 mg; 0.021 mmol) obtained in Example 4 was dissolved in pyridine (0.5 mL), acetic anhydride (0.5 mL) was added, and the mixture was stirred at room temperature for 5 hours. After concentrating the reaction solution, the obtained residue was crystallized from a mixed solvent of IPE and n-hexane to obtain 4.6 mg of Compound 3 (yield: 32%).

Compound 3:

¾-NMR (CDC1 ₃ ) δ (ppm): 0.83 (3H, t, J = 7.2 Hz), 0.84 (3H, d, J = 7.2 Hz), 0.93 C3H, d, J = 6.8 Hz), 0.99 (3H , T, J = 7.5Hz), 1.21 (2H, m), 1.56 (3H, m), 1.76 (3H, m), 1.95 (1H, m), 2.01 (3H, s), 2.04 (3H, s) , 2.13 (3H, s), 2.35 (3H, s), 2.37 (3H, s), 2.99 (IH, t, J = 5.9 Hz), 3.18 (3H, s), 3.57 (IH, m), 3.89 ( IH, d, J = 6.2Hz), 4.29 (1H, m), 5.03 (2H, s), 5.50 (1H, dd, J = 4.8, 12.1Hz), 5.87 (IH, s), 7.91 (IH, s ); FAB-MS m / z 693 [M + H] ⁺ . Example 6 Compound 4

 Compound 1 (llmg; 0.023mmol) obtained in Reference Example 1 was dissolved in methanol (lmL), sodium borohydride (4.9mg; 0.13mmol) was added, and the mixture was stirred at room temperature for 1.5 hours. . A small amount of lmol / L hydrochloric acid aqueous solution was added to the reaction solution, extracted with ethyl acetate (10 mL × 2), and the organic layer was washed with saturated saline. The organic layer was dried over anhydrous magnesium sulfate, concentrated, and the resulting residue was purified by preparative thin-layer chromatography (chloroform / methanol = 10/1) to give 6.0 mg of compound 4 (yield) 54%).

Compound 4:

-NMR (CDCI3) δ (ppm): 0.80 (3H, d, J = 6.8 Hz), 0.89 (3H, t, J = 7.3 Hz), 1.01 (3H, t, J = 7.5 Hz), 1.02 (3H, d, J = 7.5 Hz), 1.26 (2H, m), 1.54-1.64 (3H, m), 1.79 (3H, m), 2.00 (IH, m), 2.67 (IH, brs), 2.74 (IH, brs) ), 2.91 (1H, t, J = 6.4Hz), 2.98 (IH, d, J = 2.9Hz), 3.27 (IH, brs), 3.38 (3H, s), 3.47 (IH, m), 3.72 -3.89 (2H, m), 3.96 (1H, t, J = 7.5Hz), 4.18 (IH, brs), 4.88 (3H, brs), 7.41 (IH, s), 9.32 (IH, s), 13.29 (IH, s); FAB-MS m / z 507 [M + Na] ⁺ .

Example 7: Compound 6

Compound 1 (10 mg; 0.021 mmol) obtained in Reference Example 1 was dissolved in a mixed solvent of THF (1 mL) and water (0.5 mL), and 0-methylhydroxylamine hydrochloride (22 mg; 0.27 mmol) was added. The mixture was stirred at room temperature for 4 hours. After the reaction solution was diluted with ethyl acetate (10 mL), water was added for extraction, and the organic layer was washed with an aqueous solution of lm ₀ L / Lii acid. The organic layer was dried over anhydrous magnesium sulfate, concentrated, and the resulting residue was crystallized from a mixed solvent of IPE and n-hexane to obtain 5.6 mg of Compound 6 (yield: 52%).

Compound 6:

¾-NMR (CDC1 ₃ ) δ (ppm): 0.84 (3H, t, J = 7.3 Hz), 0.86 (3H, d, J = 6.8 Hz), 0.96 (3H, d, J = 6.8 Hz), 1.06 ( 3H, t, J = 7.5Hz), 1.20 (2H, m), 1.48 (IH, m), 1.58 (1H, s), 1.60 (2H, m), 1.81 (2H, m), 1.99 (2H, m) ), 2.98 (1H, brs), 3.22 (3H, s), 3.26 (IH, t, J = 6.1 Hz), 3.70 (IH, dd, J = 7.3, 14.1 Hz), 3.99 (3H, s), 4.17 (1H, td, J = 11.2, 5.1Hz), 4.34 (1H, d, J = 6.6Hz), 4.40 (IH, brt, J = 8.3Hz), 4.97 (1H, d, J = 3.5Hz), 7.83 (IH, s), 8.69 (1H, s), 11.45 (1H, s), 13.75 (IH, s); FAB-MS m / z 532 [M + Na] ⁺ , 510 [M + H] ⁺ .

Example 8: Compound 1

 Compound 1 (33.5 mg; 0.0698 mmol) obtained in Reference Example 1 was dissolved in methanol (2 mL), and a 2 mol / L trimethylsilyldiazomethane solution in n-hexane (lmL) was added. For 3.5 hours. Acetic acid (2 drops) and ethyl acetate (30 mL) were added to the reaction solution, and then water was added for extraction. The organic layer was washed with a saturated saline solution and dried over anhydrous magnesium sulfate. After evaporating the solvent, the obtained residue (45.6 mg) was purified by silica gel preparative thin-layer chromatography (n-hexane Z-ethyl acetate = lZl) to obtain 13.9 mg of compound 14 (yield 38). %).

Compound 14:

_{¾-NMR (CDC1 3, 400} MHz) δ (ppm): 12.91 (IH, s), 7.81 (IH, s), 4.97 (1H, d, J = 2.4 Hz), 4.41 (IH, m), 4.18 ( IH, m), 4.15 (IH, d, J = 6.8 Hz), 3.77 (2H, d, J = 5.6 Hz), 3.75 (3H, s), 3.72 (IH, m), 3.26 (IH, dd, J = 5.9, 7.1 Hz), 3.22 (3H, s), 2.93 (IH, brs), 2.26 (3H, s), 2.01 (IH, m), 1.92 (IH, m), 1.84 (2H, m), 1.58 (3H, m), 1.47 (IH, m), 1.20 (2H, m), 1.06 (3H, t, J = 7.6 Hz), 0.98 (3H, d, J = 6.6 Hz), 0.86 (3H, t, J = 7.3 Hz), 0.81 ( 3H, d, J = 6.8 Hz); 13 C-NMR (CDC1 3, 100 MHz) 6 (ppm): 208.0, 206.3, 164.1, 162.0, 129.2, 125.3, 117.6, 117.4- 94.5, 82.8, 69.7, 62.7, 61.9, 61.8, 59.7, 57.0, 49.9, 39.0, 37.3, 34.6, 32.0, 29.7, 20.7, 20.6, 19.1, 18.3, 14.3, 10.6; FAB-MS m / z 523 [M + H] ⁺ . Example 9: Compound 15

Reference Example 1 Compound obtained in 1 (21. ⁵ mg; 0.0448mmol) Jikurorome Yun solution (2 mL) in 4A Morekyurashi one Buss (30 mg) was added, then 4-methylmorpholine - - Okisai de (18.6 mg; 0.159 mmol) in dichloromethane (0.5 mL) and tetra (n-propyl) ammonium palmitate (3.9 mg; O.Ollmmol) in dichloromethane (0.5 mL) were added, and the mixture was stirred at room temperature for 24 hours. After the reaction solution was filtered, purification was carried out by silica gel gel column chromatography (1% methanol in chloroform solution) to obtain 1.8 mg of Compound 15 (yield: 8.4%).

Compound 15:

_{¾-NMR (CDC1 3, 500} MHz) (5 (ppm): 14.03 (IH, s), 13.01 (IH, s), 10.42 (1H, s), 8.03 (IH, s), 5.20 (IH, d, J = 3.2 Hz), 4.84 (IH, m), 4.34 (IH, d, J = 6.1 Hz), 3.77 (IH, m), 3.56 (1H, dd, J = 5.6, 7.2 Hz), 3.22 (3H, s), 3.01 (IH, d, J = 3.2 Hz), 2.68-2.63 (2H, m), 2.00-1.80 (2H, m), 1.79-1.60 (3H, m), 1.26 (2H, m), 1.08 (3H, t, J = 7.6 Hz), 0.94 (3H, d, J = 6.7 Hz), 0.90 (3H, d, J = 6.9 Hz), 0.89 (3H, t, J = 7.5 Hz); FAB-MS m / z 479 [M + H] +. Example 10: Compound 17

 Compound 12 (67.3 mg; 0.128 mmol) obtained in Reference Example 9 was dissolved in ethyl acetate (3 mL), 10% palladium-carbon (50 mg) was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 4 hours. After the catalyst was filtered off from the reaction solution, purification was performed by silica gel preparative thin-layer chromatography (mixed solvent of n-hexane and ethyl acetate) to obtain 30.3 mg of compound 17 (yield 63%).

Compound 17:

¾-NMR (CDCI3, 300 MHz) d (ppm): 7.03 (2H, d, J = 8.6 Hz), 6.85 (2H, d, J = 8.6 Hz), 6.40 (IH, m), 6.07 (IH, brs ), 5.31 (1H, m), 4.86 -4.28 (2H, m), 3.79 (3H, s), 3.61 (1H, m), 3.19 (IH, dd, J = 5.5, 10.5 Hz), 3.04 (IH, m), 2.80 (IH, d, J = 5.5 Hz), 2.76-2.49 (4H, m), 2.36 (1H, m), 2.19 (IH, brd, J = 12.5 Hz), 1.71 (IH, brd, J = 15.8 Hz), 1.49 (3H, s), 1.23 (2H, m), 1.19 (3H, d, J = 6.8 Hz), 1.14 (3H, s), 1.00 (3H, d, J = 7.2 Hz); FAB-MS m / z 528 [M + H] + .

Example 11: Compounds 18 and 19

 Compound 12 (36.6 mg; 0.0697 mmol) obtained in Reference Example 9 was dissolved in chloroform (2 mL), a small amount of p-toluenesulfonic acid monohydrate was added, and the mixture was stirred at room temperature for 5 hours. After concentrating the reaction mixture, the mixture was purified by preparative thin-layer chromatography on silica gel (mixed solvent of n-hexane and ethyl acetate) to give compound 18 (26.8 mg; yield 73%) and compound 19 ( 4.8 mg; yield 13%).

Compound 18:

_{¾-NMR (CDC1 3, 300} MHz) 6 (ppm): 7.04 (2H, d, J = 8.6 Hz), 6.84 (2H, d, J = 8.6 Hz), 6.59 (IH, d, J = 11.7 Hz) , 5.81 (IH, brs), 5.72 (IH, dd, J = 9.9, 14.9 Hz), 5.63 (IH, d, J = 11.7 Hz), 5.39 (IH, brs), 5.37 (1H, m), 5.17 ( IH, brs), 3.80 (IH, m), 3.78 (3H, s), 3.35 (1H, m), 3.24 (IH, m), 2.98-2.92 (3H, m), 2.85 (IH, dd, J = 3.9, 13.8 Hz), 2.69 (IH, m), 2.57 (IH, dd, J = 9.5, 13.8 Hz), 2.15 (IH, brd, J = 13.4 Hz), 1.52 (3H, s), 1.16 (3H, d, J = 7.2 Hz), 1.14 (3H, d, J = 7.2 Hz); FAB-MS m / z 526 [M + H] +.

Compound 19:

 ¾-NMR (CDCI3, 300 MHz) 6 (ppm): 7.00 (2H, d, J = 8.6 Hz), 6.79 (2H, d, J = 8.6.Hz), 6.57 (IH, d, J = 11.6 Hz) , 6.13 (1H, m), 6.09 (1H, brs), 5.59 (1H, d, J = 11.6 Hz), 5.31 (IH, m), 3.81 (2H, m), 3.72 (3H, s), 3.36 ( 1H, brt, J = 6.8 Hz), 2.87 (IH, m), 2.76-2.60 (4H, m), 2.09 (1H, brd, J = 10.3 Hz), 1.61 (3H, s), 1.44 (6H, s ), 1.10 (3H, d, J = 6.8 Hz); FAB-MS m / z 526 [M + H] +.

Example 12: Compound 20

From the slant culture of the filamentous fungi of the MPC1006 strain, each platinum loop was filled with 50 mL of seed medium (3% mash, 3% glucose, 10% glucose, 0.5% yeast extract, pH 6.5). And inoculated on a rotary stirrer at 28 ° C. for 5 days to obtain a seed culture. Production medium of seed culture 2.5mL of this 50 mL (2% glucose, mashed 2%, Bae script down _{0.5%, H 2 PO 4 0.5} %, Mg 3 (O 4) 2 '8H 2 O0.05%, pH6 .0) containing 300mL ^ Scos (40) were inoculated and cultured on a rotary shaker at 25 ° C for 5 days. After completion of the culture, the culture solution (2 L) was separated into bacterial cells and supernatant by suction filtration, and the culture supernatant was subjected to column chromatography using Diaion HP-20 (30 mL) previously filled with 20% methanol. After washing stepwise with 40, 50, and 60% methanol, elution was performed with 90 and 100% methanol. The eluate (150 mL) was concentrated, extracted with ethyl acetate, and the extract was separated and purified by high-performance liquid chromatography (Develosil HG-5 20x250 mm; 40-; eluted stepwise with L00% aqueous acetonitrile). 2.2 mg of compound 20 was obtained.

Compound 20:

_{¾-NMR (CDC1 3, 400} MHz) δ (ppm): 7.15 (1H, dd, J = 2.2, 15.4 Hz), 6.25 (1H, brs), 6.19 (1H, brd, J = 11.0 Hz), 6.03 ( 1H, dd, J = 2.2, 15.4 Hz), 5.30 (1H, m), 4.55 (1H, m), 3.88 (1H, dd, J = 2.4, 7.6 Hz), 3.82 (1H, brd, J = 11.0 Hz ), 3.17 (1H, dt, J = 10.0, 3.4 Hz), 3.09 (1H, m), 2.93 (1H, dd, J = 3.2, 4.9 Hz), 2.39-2.34 (1H, m), 2.20-2.08 ( 2H, m), 1.73 (3H, s), 1.61 (1H, m), 1.50-1.46 (1H, m), 1.43-1.34 (1H, m), 1.39 (3H, s), 1.33 -1.25 (1H, m), 1.21 (3H, d, J = 7.3 Hz), 0.931 (3H, d, J = 6.6 Hz), 0.926 (3H, d, J = 6.6 Hz); ¹³ C-NMR (CDCI3, 100 MHz) ( 5 (ppm): 173.6, 167.5, 150.4, 140.2, 139.5, 124.2, 122.8, 120.1, 88.3, 78.1, 73.9, 52.1, 51.7, 48.5, 39.39, 39.36, 34.2, 28.0, 25.2, 23.6, 21.4, 19.7, 15.2 , 13.8; FAB-MS m / z 418 [M + H] ⁺ .

Example 13: Compound 21

From the slant culture of the MPC1009 strain, place each loop into a 300 mL Erlenmeyer flask containing four 50 mL primary seed media (3% mashed potato, 10% glucose, 0.5% yeast extract, pH 6.5). The seeds were inoculated and cultured on a rotary stirrer at 28 ° C for 4 days to obtain a primary seed culture. 75 mL of this primary seed culture is mixed with 2.5 L of a secondary seed medium (glucose 2%, marsh potato 1%, peptone 0.5%, KH ₂ PO ₄ 0.5%, Mg PO ₄ ) ₂ '8H ₂ O 0.05%, ρΗ6.θ ) Was inoculated into a 5 L section of the yeast overnight, followed by aeration and agitation culture at 25 ° C for 24 hours. The resulting secondary-seed culture 450mL of 15L of production medium (2% glucose, mashed 2%, Bae script down _{0.5%, KH 2 PO 4 0.5} % s Mg PO 4) 2 '8H 2 O0.05%, pH6 .0) was inoculated into 2 jars of jar fermenter having a capacity of 30 L and aerated and stirred at 25 ° C for 5 days. After cultivation, The culture solution (30 L) was separated into cells and supernatant by suction filtration, and 20 L of methanol was added to the cells, followed by extraction by stirring. The extract was subjected to column chromatography with Dyaion HP-20 (1.5 L) previously filled with a 30% aqueous methanol solution, washed with a 50% aqueous methanol solution, and eluted with 100% methanol. The eluate was concentrated, extracted with ethyl acetate, and concentrated to dryness to obtain 23.7 g of bacterial cell extract. The obtained bacterial cell extract was subjected to silica gel column chromatography (a mixed solvent of black form and methanol), and a fraction containing compound 21 was collected and concentrated to dryness. The resulting residue was partitioned between a 90% aqueous methanol solution and n-hexane to remove fatty acid components. The extract (5.44 g) in a 90% aqueous methanol solution was subjected to silica gel column chromatography (a mixed solvent of n-hexane and ethyl acetate), and the fraction containing compound 21 was collected and concentrated. The obtained residue (1.24 g) was dissolved in a small amount of ether, and powdered by adding n-hexane to obtain compound 21 (343 mg).

Compound 21:

_{¾-NMR (CDC1 3, 500} MHz) δ (ppm): 7.33 (1H, dd, J = 2.5, 15.4 Hz), 6.94 (1H, brs), 6.22 (1H, d, J = 10.8 Hz), 5.91 ( 1H, dd, J = 2.1, 15.4 Hz), 5.29 (1H, brs), 4.51 (1H, m), 3.80 (1H, d, J = 10.8 Hz), 3.14 (1H, ddd, J = 3.0, 3.0, 10.3 Hz), 3.08 (1H, m), 2.89 (1H, dd, J = 3.3, 4.8 Hz), 2.23 (1H, dd, J = 9.6, 13.6 Hz), 2.11 (1H, m), 2.01 (1H, m m), 1.88 (1H, brs), 1.75 (1H, m), 1.73 (3H, s), 1.68 (1H, m), 1.63 (1H, m), 1.53 (1H, m), 1.50 (1H, m ), 1.44 (3H, s), 1.28 (1H, m), 1.20 (3H, d, J = 7.3 Hz), 0.92 (3H, d, J = 6.8 Hz), 0.91 (3H, d, J = 6.8 Hz) ^{); 13 C-NMR (CDC1} 3, 125 MHz) δ (ppm): 173.7, 167.7, 156.5, 140.1, 139.4, 124.4, 123.0, 118.8, 88.4, 69.2, 52.2, 51.9, 48.6, 42.2, 39.5, 38.1, 34.2, 25.0, 23.8, 21.3, 19.8, 17.8, 16.2, 13.9; FAB -MS mz 402 [M + H] +.

Example 14: Compound 16

From the slant culture of the filamentous fungi of the MPC1005 strain, a loop of each platinum loop was filled with four 50 mL seed culture media (3% matsushpotato, 10% glucose, 0.5% yeast extract, pH 6.5) in a 300 mL triangle. The flask was inoculated and cultured on a rotary stirrer at 28 ° C. for 5 days to obtain a seed culture. 2.5 mL of this seed culture was added to 50 ⁵ OmL production media (3% sucrose, 2% soluble starch, Inoculate a 300 mL Erlenmeyer flask containing dried yeast 0.5%, malt extract 1%, corn steep liquor (CSL) 0.5%, vegetable juice 20%, CaCO ₃ 0.5%, pH 6.0 C. The culture was performed on a rotary shaker for 5 days.

After completion of the culture, the culture solution (2.5 L) was separated into cells and supernatant by suction filtration, 6 L of methanol was added to the cells, and the mixture was stirred and extracted. The extract was concentrated under reduced pressure until the volume became about 3 L. The culture supernatant and the concentrated cell extract were combined, subjected to column chromatography on Diane HP-20 (150 mL) packed with 50% methanol, washed with 50% methanol, and then washed with 100% methanol. It was eluted at the medium. The eluate was concentrated under reduced pressure to about 200 mL, extracted with ethyl acetate (250 mL × 2), washed with saturated saline, and dried over anhydrous sodium sulfate. The residue (1.58 _g ) obtained by concentrating to dryness was dissolved in a small amount of methanol, and the column chromatography (Sephadex LH-2 °) pre-filled with chloroform-form-methanol (1: 1) (internal diameter 3.0 cm, high 20 cm), developed with a mixed solvent of chloroform and methanol (1: 1), and the eluate was fractionated by about 20 mL. Fractions containing the compound 16 were collected and concentrated to dryness under reduced pressure. The dried product was subjected to silica gel gel column chromatography (a mixed solvent of 50-m resin and chloroform) to obtain Compound 16 (I8 mg).

Compound 16:

_{¾-NMR (CDC1 3, 500MHz} ) d (ppm): 7.05 (2H, d, J = 8.5 Hz), 6.81 (2H, d, J = 8.5 Hz), 6.66 (IH, brs), 6.39 (1H, m ), 5.79 (1H, ddd, J = 1.7, 10.0, 15.1 Hz), 5.44 (1H, ddd, J = 3.9, 11.2, 15.1 Hz), 3.62 (IH, t, J = 5.9 Hz), 3.33 (IH, m), 3.32 (IH, m), 3.02 (IH, dd, J = 6.8, 12.1 Hz), 2.89 (IH, dd, J = 5.4, 10.0 Hz), 2.75 (2H, m), 2.67 (1H, dd , J = 2.1, 5.9 Hz), 2.59 (IH, d, J = 5.4 Hz), 2.25 (IH, m), 2.24 (IH, m), 2.03 (IH, dt, J = 13.8, 11.6 Hz), 1.88 (3H, d, J = 0.7 Hz), 1.17 (3H, s), 1.15 (3H, d, J = 6.6 Hz), 1.11 (3H, d, J = 7.3 Hz); ¹³ C-NMR (CDCI3, 125 MHz) ό "(ppm): 206.2, 171.9, 169.1, 155.1, 143.1, 135.1, 132.7, 130.8, 130.6, 128.7, 126.1, 115.8, 84.9, 60.4, 57.2, 54.4, 49.5, 47.0, 43.9, 39.9, 39.8, 36.8, 36.1, 19.7, 17.4, 13.0, 12.8; FAB -MS m / z 480 [M + H] +.

Reference Example 1: Compound 1 (UCS15A / Luminacin C2 / SP4228A) Compound 1 was produced by the method described in The Journal of Antibiotics, vol. 53, p. 579 (2000) and JP-A-58-116686. It was obtained by culturing actinomycetes belonging to the genus Streptoy and isolating and purifying it from the culture broth.

Compound 1 (UCS15A / Luminacin C2 / SI-4228A):

_{¾-NMR (CDC1 3, 300} MHz) δ (ppm): 14.16 (IH, s), 12.93 (IH, s), 10.35 (IH, s), 8.01 (IH, s), 4.85 (1H, d, J = 3.2 Hz), 4.36 (IH, dd, J = 8.1, 9.6 Hz), 4.27 (IH, d, J = 7.0 Hz), 4.11 (1H, ddd, J = 4.6, 11.4, 11.4 Hz), 3.66 (IH , m), 3.53 (IH, d, J = 3.2 Hz), 3.19 (IH, t, J = 6.2 Hz), 3.14 (3H, s), 1.85 -1.98 (2H, m), 1.69-1.78 (2H, m), 1.40-1.58 (3H, m), 1.08-1.19 (2H, m), 0.98 (3H, t, J = 7.5 Hz), 0.91 (3H, d, J = 6.6 Hz), 0.79 (3H, t , / = 7.3 Hz), 0.77 (3H, d, J = 6.8 Hz); FAB-MS mlz 479 [MH] ".

Reference Example 2: Compound 5

 Compound 5 can be obtained according to the method described in JP-A-62-294619, but can also be obtained by the following method.

Reference Example 1 Compound obtained in 1; dissolved (Llmg 0.023 mmol) in a mixed solvent of THF (LML) and water (0.5 mL) and methanol Ichiru (1 drop), sulfate methylhydrazine ⁽³ 0 mg, 0. ² lmmol) and stirred at room temperature for 4 hours. The reaction mixture was diluted with ethyl acetate (lOmL) and extracted, and the organic layer was washed with water and an aqueous solution of lmol / L ^ acid. The organic layer was dried over anhydrous magnesium sulfate and concentrated, and the obtained residue was crystallized from a mixed solvent of IPE and n-hexane to obtain 5.9 mg of Compound 5 (yield 50%).

Compound 5:

¾-NMR (CDCI3) (J (ppm): 0.84 (3H, t, J = 7.2 Hz), 0.86 (3H, d, J = 6.8 Hz), 0.96 (3H, d, J = 6.6 Hz), 1.06 ( 3H, t, J = 7.5Hz), 1.21 (2H, m), 1.42-1.62 (3H, m), 1.82 (2H, m), 2.01 (2H, m), 3.00 (3H, s), 3.22 (3H , s), 3.26 (IH, t, J = 6.2 Hz), 3.70 (IH, m), 4.16 (1H, m), 4.35 (IH, d, J = 6.6 Hz), 4.40 (IH, m), 4.98 (IH, s), 6.72 (IH, s), 7.74 (IH, s), 8.21 (IH, s), 13.00 (1H, brs), 13.71 (IH, s); FAB-MS mlz 477 [M-CH ₃ NH ₂ ] +, 507 [MH] \ Compounds 7, 8, and 9 are published in the journal 'OB' American 'Chemical Society (J. Am. Chem. Soc.), Vol. 122, p. 3071 (2000). Reference Example 3: Compound 7

 Resorcinol (600 mg; 5.45 mmol) and sorbic acid (600 mg; 5.36 mmol) were dissolved in boron trifluoride etherate (10 mL) and stirred at 120 ° C. for 15 minutes. A small amount of water was added to the reaction solution while cooling with ice, and the mixture was extracted with ethyl acetate (50 mL x 2). After evaporating the solvent from the organic layer under reduced pressure, the residue was dissolved in a mixed solvent of THF (50 mL) and water (50 mL), and the mixture was heated under reflux for 30 minutes. After the reaction solution was concentrated and THF was distilled off, the mixture was extracted with ethyl acetate (50 mL × 2), and the organic layer was dried over anhydrous magnesium sulfate. The organic layer was concentrated, and the obtained residue (1.54 g) was purified by silica gel column chromatography (75 g; n-hexane / ethyl acetate = 3: 1) to obtain 566 mg of compound 7 (yield: 60%). . Compound 7:

¾-NMR (CDC1 ₃ ) 5 (ppm): 1.92 (3H, d, J = 5.1 Hz), 5.57 (1H, brs), 6.32-6.40 (4H, m), 6.91 (1H, d, = 14.7 Hz) , 7.48 (1H, dd, J = 9.9, 14.7Hz), 7.71 (1H, d, J = 9.2Hz), 13.41 (1H, s); FAB-MS mlz 205 [M + H] ⁺ .

Reference Example 4: Compound 8

 736 mg of Compound 8 was obtained from 2-methylresorcinol (500 mg; 4.03 mmol) and sorbic acid (450 mg; 4.01 mmol) in the same manner as in Reference Example 3 (yield 84%). Compound 8:

 ¾-NMR (CDCl,) δ (ppm): 1.90 (3H, d, J = 5.5Hz), 2.14 (3H, s), 6.00 (1H, brs),

6.23- 6.35 (2H, m), 6.39 (1H, d, J = 9.0Hz), 6.92 (1H, d, J = 15.0Hz), 7.46 (1H, dd, J = 9.9, 15.0Hz), 7.58 (1H , D, J = 9.0Hz), 13.73 (1H, s); FAB-MS mlz 219 [M + H] +.

Reference Example 5: Compound 9

 From 2,4-dimethylresorcinol (1.08 g; 7.83 mmol) and sorbic acid (950 mg; 8.47 mmol), 971 mg of compound 9 was obtained in the same manner as in Reference Example 3 (yield 53%). Compound 9:

 ¾-NMR (CDCI3) 51.91 (3H, d, J = 5.7 Hz), 2.15 (3H, s), 2.22 (3H, s), 5.32 (1H, s),

6.24- 6.40 (2H, m), 6.95 (1H, d, J = 14.9Hz), 7.45 (1H, s), 7.46 (1H, dd, J = 10.1, 14.9Hz), 13.60 (1H, s); FAB-MS m / z 233 [M + H] ⁺ .

Reference Example 6: Compound 10

 Compound 10 can be obtained by the method described in Tetrahedron, vol. 31, p. 1593 (1975), but can also be obtained by methylation of compound 7. Compound 7 (424 mg; 2.08 mmol) obtained in Reference Example 3 was dissolved in 30 mL of acetone, and calcium carbonate (650 mg; 4.70 mmol) and methyl iodide (0.260 mL; 4.18 mmol) were added, followed by heating under reflux for 1 hour. . The reaction solution was allowed to cool at room temperature, diluted with ethyl acetate (50 mL), and extracted with lmol / L aqueous acid solution. The organic layer was washed with brine, dried over anhydrous magnesium sulfate, and concentrated. The residue (410 mg) obtained was purified by silica gel column chromatography (30 mL; n-hexane Z ethyl acetate = 4Zl) to give the compound. 113 mg of 10 were obtained (yield 25%).

Compound 10:

_{¾-NMR (CDC1 3) S} (pp): 1.91 (3Η, d, J = 5.0Hz), 3.85 (3H, s), 6.26-6.40 (2H, m), 6.43-6.47 (2H, m), 6.92 (1H, d, J = 14.9Hz), 7.48 (1H, dd, J = 9.9, 14.9Hz), 7.71 (1H, d, J = 9.5Hz), 13.50 (1H, s); FAB-MS m / z 219 [M + H] ⁺

Reference Example 7: Compound 11

Compound 7 (I37mg; 0.672mraol) obtained in Reference Example 3 was dissolved in 25mL of acetone, and calcium carbonate (l.50g; 10.85mmol) and methyl iodide (l.50mL; 24.1mmol) were added. Heated to reflux. The same post-treatment as in Reference Example 6 was performed to obtain 122 _{mg of} Compound 11 (yield: 78%).

Compound 11:

-NMR (CDCI3) 6 (ppm): 1.85 (3H, d, J = 6.2Hz), 3.83 (3H, s), 3.84 (3H, s), 6.12-6.32 (2H, m), 6.46 (1H , d, J = 2.2Hz), 6.51 (1H, dd, J = 2.2, 8.6Hz), 6.81 (1H, d, J = 15.2Hz), 7.25 (1H, dd, J = 11.0, 15.0Hz), 7.66 (1H, d, J = 8.6 Hz); FAB-MS m / z 233 [M + H] ⁺ . Reference Example 8: Compound 13 (Cytochalasin E)

Compound 13 was published in the journal 'Op' Chemical Society, Parkin Transa ύ

Cushion 1 (J. Chem. Soc., Perkin Trans. 1), 541 (1982), Agricultural and Biological, Chemistry (Agric. Biol. Chem.), 53, 1699 (1989 According to the method described in the above, etc., a filamentous fungus having the ability to produce the compound was cultured and isolated and purified from a culture solution.

Compound 13 (Cytochalasin E):

_{-NMR (CDC1 3, 300 MHz)} d (ppm): 7.27-7.37 (3H, m), 7.15 (2H, d, J = 7.0 Hz), 6.53 (1H, d, / = 12.5 Hz), 6.00 (1H , brs), 5.89 (1H, dd, J = 8.1, 15.0 Hz), 5.63 (1H, d, J = 12.5 Hz), 5.23 (1H, m), 3.71 (1H, m), 3.02 (1H, m) , 2.94 (1H, m), 2.89 (1H, dd, J = 4.2, 13.9 Hz), 2.50-2.70 (4H, m), 2.31 (1H, m), 2.15 (1H, brd, J = 15.0 Hz), 1.50 (3H, s), 1.25 (3H, s), 1.16 (3H, d, J = 6.1 Hz), 1.12 (3H, d, / = 6.8 Hz); FAB-MS m / z 496 [M + H] ⁺ Reference Example 9: Compound 1 2 (Mer-WF1726)

 Compound 12 can be obtained according to the method described in JP-A-10-114776.

Compound 1 2 (Mer-WF1726):

NMR-NMR (CDCI3, 500 MHz) δ (ppm): 7.06 (2H, d, J = 8.5 Hz), 6.86 (2H, d, J = 8.5 Hz), 6.51 (1H, d, J = 11.6 Hz), 6.16 (1H, brs), 5.90 (1H, dd, J = 9.3, 14.9 Hz), 5.61 (1H, d, J = 11.6 Hz), 5.23 (1H, ddd, J = 3.8, 10.9, 14.9 Hz), 3.79 (3H, s), 3.68 (1H, m), 3.00 (1H, dd. J = 2.6, 5.1 Hz), 2.94 (1H, m), 2.82 (1H, dd, J = 4.5, 13.7 Hz), 2.69- 2.60 (4H, m), 2.29 (1H, dq, J = 12.5, 7.3 Hz), 2.15 (1H, brd, J = 13.8 Hz), 1.49 (3H, s), 1.25 (3H, s), 1.16 (3H , d, J = 6.8 Hz) , 1.10 (3H, d, J = 7.3 Hz); 13 C-NMR (CDC1 3, 125 MHz) 6 (ppm): 211.8, 170.0, 158.9, 149.4, 142.2, 131.6, 130.6 , 128.5, 127.9, 120.5, 114.4, 87.1, 77.3, 60.7, 57.3, 55.3, 53.8, 48.0, 45.9, 44.1, 40.9, 39.1, 35.9, 24.4, 20.1, 19.7, 13.3; FAB-MS m / z 526 [M + H] ⁺ .

Reference Example 10: Compound 26 (Aspochalasin C), Compound 27 (Aspochalasin D) and Compound 28 (Aspochalasin E)

Compound 26 (Aspochalasin C), Compound 27 (Aspochalasin D) and Compound 28 (Aspochalasin E) should be cultured and isolated and purified from MPC1009 filamentous fungi by the method described in Example 13. Was obtained. Also, Helvetica Chim. Acta, Vol. 62, p. 1501 (1979), Journal of Antibiotics (J. Antibiot.), Vol. 46, p. 679 ( 1993), and the like, by culturing a filamentous fungus capable of producing the compound and isolating and purifying the same from a culture solution.

Compound 26 (Aspochalasin C):

_{¾-NMR (CDC1 3, 400} MHz) δ (ρρτα): 7.32 (1Η, dd, J = 0.7, 16.3 Hz), 6.26 (1H, dd, J = 8.3, 16.3 Hz), 6.07 (1H, brs), 5.99 (1H, brd, J = 11.2 Hz), 5.41 (1H, m), 3.95 (1H, t, J = 8.3 Hz), 3.49 (1H, m), 3.10 (1H, m), 3.02 (1H, dd , J = 3.7, 5.4 Hz), 2.82 (1H, brd, J = 11.0 Hz), 2.46 (2H, m), 2.02 (1H, m), 1.93 (1H, m), 1.85 (1H, ddd, J = 2.7, 13.2, 13.2 Hz), 1.77 (3H, d, J = 1.5 Hz), 1.55 (1H, m), 1.41 (3H, d, J = 1.5 Hz), 1.24 (2H, m), 1.23 (3H, d, J = 7.3 Hz), 0.91 (3H, d, J = 6.6 Hz), 0.90 (3H, d, J = 6.6 Hz); 13 C-NMR (CDC1 3, 100 MHz) d (ppm): 198.2, 174.4, 140.8, 138.3 (2C), 130.3, 125.8, 125.5, 77.3, 75.1, 68.0, 51.3, 49.7, 48.4, 43.5, 36.4, 36.3, 35.2, 25.2, 23.7, 21.4, 20.0, 19.9, 13.7; FAB-MS mlz Ql [M + H] +.

Compound 27 (Aspochalasin D):

_{¾-NMR (CDC1 3, 300} MHz) d (ppm): 7.14 (1H, d, J = 16.7 Hz), 6.14 (1H, dd, J = 5.0, 16.5 Hz), 6.33 (1H, brs), 5.94 ( 1H, d, J = 10.8 Hz), 5.42 (1H, brs), 4.58 (1H, brs), 3.78 (1H, brs), 3.15 (1H, m), 3.00 (1H, dd, J = 2.8, 5.7 Hz) ), 2.90 (1H, d, J = 10.8 Hz), 2.48 (1H, m), 2.16 (2H, m), 2.07 (1H, m), 1.75 (3H, s), 1.50 (2H, m), 1.30 (3H, s), 1.22 (3H, d, J = 7.3 Hz), 1.21 (1H, m), 0.90 (3H, d, J = 6.4 Hz), 0.88 (3H, d, J = 6.4 Hz); ¹³ C-NMR (CDCI3, 75 MHz) (5 (ppm): 197.6, 174.9, 141.5, 140.4, 137.4, 129.6, 125.7, 124.2, 79.2, 75.6, 68.0, 51.0, 49.6, 48.3, 43.6, 39.5, 35.1, 29.3 , 25.1, 23.6, 21.5, 19.9, 15.6, 13.5; FAB-MS mlz 402 [M + H] ⁺ . Compound 28 (Aspochalasin E):

¾-image R (DMSO-d ₆ , 500 MHz) d (ppm): 8.10 (1H, s), 6.02 (1H, d, J = 10.9 Hz), 5.33 (1H, brs), 4.41 (1H, d, J = 5.9 Hz), 4.37 (1H, d, J = 4.0 Hz), 4.35 (1H, brs), 3.90 (1H, d, J = 17.6 Hz), 3.60 (1H, brs), 3.24 (1H, brs) , 3.17 (1H, brs), 3.07 (2H, m), 2.48 -2.40 (2H, m), 2.00-1.94 (2H, m), 1.76 (1H, dd, J = 5.2, 17.6 Hz), 1.70 (3H, s), 1.60-1.44 (2H, m), 1.38 (3H, d, J = 1.1 Hz), 1.31 (1H, m), 1.16 (3H, d, J = 6.9 Hz), 1.09-0.98 (2H, m), 0.84 (3H, d, J = 6.6 Hz), 0.83 (3H, d, J = 6.5 Hz); FAB-MS m / z 420 [M + H] ⁺ .

Reference Example 11 1: Compound 22 and Compound 25 (Aspochalasin B)

 Compound 27 (31.7 mg; 0.0791 mmol) obtained in Reference Example 10 was dissolved in dichloromethane (5 mL), manganese dioxide (250 mg; 2.44 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered using celite, and the filtrate was concentrated. The residue was purified by silica gel preparative thin-layer chromatography (a mixed solvent of chloroform and methanol) to give Compound 25 (13.6 mg; yield 43%). %) And compound 22 (5.1 mg; yield 16%).

The compound 2 5 (Aspochalasin B) is Herube Chika-Himika '§ click evening (Helv. Chim. Acta), 6 2 Certificates, according to the method described in 1501 pages ^{(197, 1997),} the production of the compound It can also be obtained by culturing a functional filamentous fungus and isolating and purifying it from a culture solution. .

Compound 25 (Aspochalasin B):

_{¾-NMR (CDC1 3, 300} MHz) δ (ppm): 8.28 (1H, d, J = 16.5 Hz), 6.52 ΠΗ, d, J = 16.5 Hz), 6.33 (1H, d, J = 11.2 Hz), 5.99 (1H, brs), 5.44 (1H, brs), 4.82 (1H, brd, J = 6.1 Hz), 3.59 (1H, brs), 3.19 (1H, t, J = 4.6 Hz), 3.11 (1H, m ), 2.74 (1H, brd, J = 11.4 Hz), 2.62 (1H, m), 2.38 (2H, m), 1.86 (1H, m), 1.79 (3H, brs), 1.57 (3H, m), 1.24 (3H, d, J = 7.0 Hz), 1.22 (3H, s), 0.92 (3H, d, J = 5.9 Hz), 0.90 (3H, d, J = 6.2 Hz); 13 C-NMR (CDC1 3, 75 MHz) (5 (ppm): 205.1, 195.5, 173.2, 141.6, 138.8, 136.4, 126.5 (2C), 124.7, 74.7, 68.9, 51.7, 48.2, 47.5, 41.6, 39.7, 34.8, 32.4, 25.2, 23.7, 21.1, 20.2, 15.5, 13.8; FAB-MS m / z 400 [M + H] ⁺ .

Compound 22:

¾-NMR (CDCI3, 300 MHz) δ (ppm): 9.77 (1H, brs), 9.75 (1H, d, J = 7.7 Hz), 7.76 (1H, d, J = 15.8 Hz), 6.78 (1H, dd) , J = 7.7, 15.8 Hz), 6.12 (1H, brs), 5.76 (1H, d, / = 10.3 Hz), 5.34 (1H, brs), 3.23 (1H, d, J = 9.9 Hz), 3.16 (1H , m), 2.74 (1H, t, J = 4.8 Hz), 2.54 (3H, m), 2.33 (2H, m), 1.76 (3H, brs), 1.54 (IH, m), 1.50 (3H, s), 1.34 (1H, m), 1.21 (3H, d, J = 7.3 Hz), 0.92 (6H, d, J = 6.6 Hz); FAB-MS mlz 400 [M + H] ⁺ .

Reference Example 12 2: Compound 24 (Aspochalasin A)

Compound 25 (28.9 mg; 0.0724 mmol) obtained in Reference Example 11 was dissolved in pyridine (0.5 mL), and triethylamine (0.5 mL) was added, followed by stirring at room temperature for 19 hours. The reaction solution was poured into water, extracted with chloroform (20 mL), and washed with 0.5 mol / Li aqueous acid. The product was purified by preparative thin-layer chromatography on a silica gel (prepared by chromatography on a gel) to obtain Compound 24 (21.2 mg; yield ⁷³ %).

 In addition, compound 24 is a filamentous fungus capable of producing the compound according to the method described in Helvetica Chimica Acta, Vol. 62, p. 1501 (1979). It can also be obtained by culturing and isolating and purifying from a culture solution.

Compound 24:

_{¾-NMR (CDC1 3, 300} MHz) δ (ppm): 6.73 (IH, brs), 6.22 (IH, d, J = 11.0 Hz), 5.32 (IH, brs), 3.94 (IH, dd, J = 12.8 , 19.4 Hz), 3.67 (1H, m), 3.14 -3.07 (2H, m), 2.94 (1H, brd, J = 9.5 Hz), 2.79 (IH, brt, J = 12.8 Hz), 2.55-2.40 (3H , M), 2.38-2.02 (2H, m), 1.99 (IH, m), 1.72 (3H, s), 1.56 (IH, m), 1.48 (3H, s), 1.19 (3H, J = 6.6 Hz) , 1.16 (2H, m), 0.94 (6H, d, J = 6.6 Hz); 13 C-NMR (CDC1 3, 75MHz) δ (ppm): 208.7, 202.6, 197.6, 175.5, 140.0, 135.7, 125.1, 124.8 , 66.3, 52.1, 50.7, 48.7, 43.4, 38.8, 36.6, 35.3, 31.4, 31.2, 25.0, 23.5, 21.4, 19.9, 14.8, 13.4; FAB -MS m / z 400 [M + H] +.

Reference Example 13: Compound 23

 Compound 25 (216 mg; 0.541 mmol) obtained in Reference Example 11 was dissolved in ethyl acetate (30 mL), 10% palladium on carbon (150 mg) was added, and the mixture was stirred under a hydrogen atmosphere at room temperature for 4 hours. After filtering off the catalyst, purification was performed by preparative thin-layer chromatography on silica gel (mixed solvent of chloroform and methanol) to obtain 92.3 mg of compound 23 (yield 43%).

Compound 23:

_{¾-NMR CCDC1 3, 300 MHz} ) (5 (ppm): 7.01 (IH, brs), 6.00 (IH, d, J = 10.6 Hz), 4.17 (IH, brs), 3.40 (1H, m), 2.89-2.76 (3H, m), 2.42-1.76 (7H, m), 1.64-1.44 (2H, m), 1.32 (3H, d, J = 1.1 Hz), 1.25 -1.03 (2H, m), 0.92-0.85 (15H, m); FAB-MS m / z 402 [M + H] ⁺ .

 '' Piτ Noh calf for industrial turtle II

 The present invention provides an SH3 domain binding inhibitor comprising a non-peptidic compound or a pharmacologically acceptable salt thereof as an active ingredient. The present invention also provides a compound useful as an SH3 domain binding inhibitor or a pharmacologically acceptable salt thereof.

Claims

The scope of the claims  1. An SH3 domain binding inhibitor comprising, as an active ingredient, a non-peptide compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof.  2. The SH3 domain binding inhibitor according to claim 1, wherein the non-peptide compound is a low-molecular compound having a molecular weight of less than 750.  3. A non-peptide compound having SH3 domain binding inhibitory activity is represented by the general formula (I)

(Wherein ^,, R ^3a , R ^3b and R ⁴ are the same or different and each represents a hydrogen atom, hydroxy, carboxy, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkoxy, or substituted or unsubstituted lower alkyl) R ^2a and R ^2b are the same or different and represent a hydrogen atom, a substituted or unsubstituted lower alkyl or a substituted or unsubstituted or substituted alkanoyloxy, or R ^3a and R ^3b each represent an oxygen atom, Represents an unsubstituted lower alkenyl, and R ^5a and are the same or different and are each a hydrogen atom, hydroxy, carboxy, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkoxy, substituted or unsubstituted lower alkenyl, Substituted or unsubstituted lower alkanoyloxy, substituted or unsubstituted lower alkenoyloxy or substituted Or properly are a lower alkanoyloxy Noi Rua iminocarbonyl O carboxymethyl unsubstituted, or R ^5a and R ⁵¹³ together represent an oxygen atom, X represents an oxygen atom or a CH ₂ - compound you express at representative) 3. The SH3 domain binding inhibitor according to claim 1 or 2, wherein 4. A non-peptide compound having SH3 domain binding inhibitory activity is represented by the general formula (II)

(In the formula, R ⁶ represents a hydrogen atom, substituted or unsubstituted lower alkyl or substituted or unsubstituted lower alkenyl, and R ⁷ and R ⁹ are the same or different and represent a hydrogen atom, formyl, substituted or unsubstituted Represents lower alkyl, substituted or unsubstituted lower alkanol or substituted or unsubstituted aralkyl, and R ^1Q and R ¹¹ are the same or different and are a hydrogen atom, halogen, hydroxy, carboxy, substituted or unsubstituted Lower alkyl, substituted or unsubstituted lower alkoxy, substituted or unsubstituted lower alkoxy, formyl, cyano, nitro, amino, mono- or di-lower alkylamino, substituted or unsubstituted lower alkylamino or substituted or unsubstituted Represents a substituted lower alkoxycarbonyl) SH3 domain-binding inhibitor range 1 or 2, wherein the billing.  5. The SH3 domain binding inhibitor according to claim 1, wherein the non-peptide compound having SH3 domain binding inhibitory activity is a cytochalasin.  6. A non-peptide compound having SH3 domain binding inhibitory activity is represented by the general formula (Ilia)

(In the formula, R ^{12 represents a} substituted or unsubstituted lower alkyl, a substituted or unsubstituted aryl or a substituted or unsubstituted heteroaryl, Q ¹ represents a single bond or an oxygen atom, and-represents a single bond. It represents a bond or a double bond, between the carbon atoms adjacent to Q ² - two When represents a double bond, = Q ² — represents = C (CH ₃ ) —, and when Nini between Q ² and an adjacent carbon atom represents a single bond, — Q ² — represents —C (OH) ( _3. The SH3 domain binding inhibitor according to claim 1 or 2, which is a compound represented by the following formula:  7. A non-peptide compound having SH3 domain binding inhibitory activity is represented by the general formula (lift)

 (Illb) Wherein, R ^12b are as defined above R ^12a, Q ³ and Q ⁵ are the same or different, it represents a single binding or an oxygen atom,: represents a single bond or a double bond, a Q ⁴ Nini represents = C (CH ₃ ) —, —C (= CH ₂ ) —, one CH (CH ₃ ) — or one C (CH ₃ ) =, and R ^12c and R ^12h are the same or different and are hydrogen Represents an atom or hydroxy, R ^12d and R ^12e are the same or different and represent a hydrogen atom or methyl, R ^12f and R ^12g represent formyl, or R ^12f and R ^12g together , ---- CR ^12h R ^12g and - part of CHR ^12e R ^{12f is, - (. CH) -CR 12h} -C (OH) one ^{C (= 0) - CHR 12e} - or - -CR ^12h -AB- CHR ^12e- (wherein A and B are the same or different and represent —CH (OH) —, one CH ₂ — or one C (20) —). 3. The SH3 domain binding inhibitor according to claim 1 or 2. 8. X is an oxygen atom, R, R ^3a , R ⁴ and 1 are hydrogen atoms, R ¹ and R ^3b are the same or different and are hydroxy, carboxy, substituted or unsubstituted lower alkoxy or substituted or unsubstituted A substituted lower alkanoyloxy, R ^2b is a substituted or unsubstituted lower alkyl, and R ^5a is a group represented by the general formula (IV)

Wherein R ^5e represents a substituted or unsubstituted lower alkyl, and R ^5d and R ⁵ are the same or different and represent a hydrogen atom, hydroxy or a substituted or unsubstituted lower alkoxy, or ^5d and R ^5e together represent an oxygen atom, R ^5f and R ⁵¹¹ represent a hydrogen atom, formyl, substituted or unsubstituted lower alkyl or substituted or unsubstituted lower alkanol, and R ^5g is formyl , -CH = NQ (wherein Q is substituted or unsubstituted lower alkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted lower alkylamino, substituted or unsubstituted Represents substituted aryl or substituted or unsubstituted arylsulfonylamino), substituted or unsubstituted lower alkyl or substituted or unsubstituted It represents lower Arukanoiru, R ⁵ⁱ and R ^5j are the same or different, represent a hydrogen atom, or a hydroxy or substituted or unsubstituted lower alkoxy, or taken R ⁵ⁱ and R ^5j gar cord oxygen atom 4. The SH3 domain binding inhibitor according to claim 3, wherein  9. General formula (Va)

 (Va) (Wherein, 2 represents a single bond or a double bond, and R ^12a , Q ¹ and Q ² have the same meanings as described above) or a pharmaceutically acceptable salt thereof. 10. General formula (Vb)

[Wherein, 2 represents a single bond or a double bond, and R ^12b , R ¹² \ R ^12d , R ^12e , R ^12h , Q ^5, and--_ Q ⁴ __ _-are as defined above. ■ CR ¹ -D— CHR ^12e — ™ CR ^12h -C (OH) (CH ₃ )-C (= O)-CHR ^12e -or two CR ^12h —A— B— CHR ^12e — (wherein A and B are as defined above. Or a pharmacologically acceptable salt thereof.  11. General formula (VI)

[Wherein, R ^3A and R ^3B are the same or different and each represent a hydrogen atom, hydroxy, hydroxyl, substituted or unsubstituted lower alkoxy or substituted or unsubstituted lower alkanoyloxy, or R ^3A And R ^3B represent an oxygen atom, represents a substituted or unsubstituted lower alkyl, and RR ^5d , R ^5e , R ^5f , R, R ⁵ⁱ and R ^5j are each as defined above. ^R5G is formyl, hydroxymethyl, substituted or unsubstituted lower alkoxymethyl, substituted or unsubstituted lower alkanoyloxymethyl, substituted or unsubstituted lower alkenylmethyl, or CH = NQ ^A (Wherein Q ^A represents a substituted or unsubstituted lower alkoxy, a substituted or unsubstituted aryloxy or a substituted or unsubstituted aralkyloxy) However, when R ^5G is formyl and one of R ^3A or R ^3B is a hydrogen atom, the other of R ^3A or R ^3B is not hydroxy] or a pharmaceutically acceptable compound thereof. salt.  12. A medicament comprising the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof as an active ingredient.  13. A medicament comprising the compound according to claim 11 or a pharmacologically acceptable salt thereof as an active ingredient.  14. An SH3 domain binding inhibitor comprising the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof as an active ingredient.  15. An SH3 domain binding inhibitor comprising the compound according to claim 11 or a pharmacologically acceptable salt thereof as an active ingredient.  16. The SH3 domain binding inhibitor according to any one of claims 1 to 8, 14, and 15, wherein the SH3 domain binding is a binding between a protein having an SH3 domain and a protein having a proline-rich sequence.  17. The SH3 domain binding inhibitor according to claim 16, wherein the protein having an SH3 domain and the protein having a Z or proline rich sequence are proteins derived from a virus.  18. The SH3 domain binding inhibitor according to claim 17, wherein the virus-derived protein is a protein derived from a retrovirus, a hepatitis virus, or a virus. 19. Proteins with SH3 domain are Src, Yes, Fgr, Hck, Lck, Abl, Fyn, Lyn ヽ Blk ヽ Yrk, Ras-GAP ヽ PLC, PI3K ヽ Tec, Txk / Rlk ヽ Tsk / Emt / Itk _{N Btk, Crk, Grb2 s Neks Vav} , STAT, cortactin, p40-phox, p67-phox, p47-phox, TCR Sig null molecules (TCRsm) or IL-2R of ^ chain or claims 16, wherein the a § chain SH3 domain binding inhibitor. 20. Proteins having a proline-rich sequence are HIV-lNef, p22-phox, p47-phox _N Sam68, Sosl, Dynamic c-Cbl, ZOl ヽ pX ORF I ヽ LHDAg ヽ NS5A ヽ pORF3, ICP10, 20. The SH3 domain binding inhibitor according to claim 16 or 19, which is LMP2A, Tip or Tio.  21. The binding of the protein having the SH3 domain to the protein having the proline-rich sequence is Grb2 and Sosl, Fyn and Sam68, Src and Sam68, PLC and Sam68, Grb2 and Sam68, Lyn and HIV-Nef ヽ Hck and HIV- lNef ヽ TCRsm and HIV-lNef ヽ p47-phox and p22-phox, p67-phox and p47-phox, Lyn and Dynami Cortactin and Z01, Lyn and c-Cbl, IL-2R /? or a-2 and pX ORF I, Grb2 and NS5A, Src and pORF3, Hck and pORF3, Fyn and pORF3, PI3K and pORF3, PLC and pORF3, Grb2 and pORF3, Grb2 and ICP10, Lyn and LMP2A ヽ Lck and Tip, Lyn and Ήο, Hck and Tio 17. The SH3 domain binding inhibitor according to claim 16, which is a bond between Lck and Tio, Src and Ήο, Fyn and Tio or Yes and Tio.  22.The binding of the protein having the SH3 domain and the protein having the proline-rich sequence is Grb2 and Sosl, Fyn and Sam68, Src and Sam68, PLC and Sam68, Grb2 and Sam68, Lyn and HIV-lNef or Cortactin and ZOl. 17. The SH3 domain binding inhibitor according to claim 16, which is binding.  23. ^^ used for the production of the compound according to claim 9 or 10 And filamentous fungi (J / arwfes filamentous fungus) MPC1005 (Accession number: PERM BP-7980), Aspergillus sp. (Aipergi ws sp.) MPC1006 (Accession number: PERM BP-7899) and Aspergillus species (Α g iw sp.) A microorganism selected from the group consisting of MPC1009 (Accession number: FERM BP-7900).  24. An antitumor agent comprising the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof as an active ingredient.  25. An antitumor agent comprising the compound according to claim 11 or a pharmacologically acceptable salt thereof as an active ingredient.  26. A therapeutic agent for an allergic disease, comprising the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof as an active ingredient. 27. A therapeutic agent for an allergic disease, comprising the compound according to claim 11 or a pharmacologically acceptable salt thereof as an active ingredient. 28. A therapeutic agent for a viral disease, comprising the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof as an active ingredient.  29. A therapeutic agent for a viral disease, comprising the compound according to claim 11 or a pharmacologically acceptable salt thereof as an active ingredient.  30. An AIDS therapeutic agent comprising the compound according to claim 9 or 10 or a pharmaceutically acceptable salt thereof as an active ingredient.  31. An AIDS therapeutic agent comprising the compound according to claim 11 or a pharmacologically acceptable salt thereof as an active ingredient.  32. Use of the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof for the production of an SH3 domain binding inhibitor.  33. Use of the compound according to claim 11 or a pharmaceutically acceptable salt thereof for the production of an SH3 domain binding inhibitor.  34. Use of the compound according to claim 9 or 10 or a pharmaceutically acceptable salt thereof for the manufacture of an antitumor agent.  35. Use of the compound according to claim 11 or a pharmaceutically acceptable salt thereof for the manufacture of an antitumor agent.  36. Use of the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof for the manufacture of a therapeutic agent for an allergic disease.  37. Use of the compound according to claim 11 or a pharmacologically acceptable salt thereof for the manufacture of a therapeutic agent for an allergic disease.  38. Use of the compound according to claim 9 or 10 or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for a viral disease.  39. Use of the compound according to claim 11 or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for a viral disease.  40. Use of the compound according to claim 9 or 10 or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for AIDS. 41. The compound according to claim 11 for producing an AIDS therapeutic agent or the pharmacology thereof. Use of chemically acceptable salts.  42. A method for treating and / or preventing or treating various diseases involving SH3 domain binding, which comprises administering an effective amount of the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof.  43. A method for treating and / or preventing various diseases involving SH3 domain binding, which comprises administering an effective amount of the compound according to claim 11 or a pharmacologically acceptable salt thereof.  44. A method for treating a malignant tumor, comprising administering an effective amount of the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof.  45. A method for treating a malignant tumor, which comprises administering an effective amount of the compound according to claim 11 or a pharmacologically acceptable salt thereof.  46. A method for treating and / or preventing an allergic disease, which comprises administering an effective amount of the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof.  47. A method for treating and / or preventing or treating allergic diseases, which comprises administering an effective amount of the compound according to claim 11 or a pharmacologically acceptable salt thereof.  48. A method for treating a viral disease, which comprises administering an effective amount of the compound according to claim 9 or 10 or a pharmacologically acceptable salt thereof.  49. A method for treating a viral disease, which comprises administering an effective amount of the compound according to claim 11 or a pharmacologically acceptable salt thereof.  50. A method for treating AIDS, which comprises administering an effective amount of the compound according to claim 9 or 10 or a pharmaceutically acceptable salt thereof.  51. A method for treating AIDS, which comprises administering an effective amount of the compound according to claim 11 or a pharmacologically acceptable salt thereof.  52. A method for treating AIDS, which comprises administering an effective amount of a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof. 53. Compound having SH3 domain binding inhibitory activity or its pharmacologically acceptable A method for treating a viral disease, which comprises administering an effective amount of a salt to be treated.  54. An AIDS therapeutic agent comprising, as an active ingredient, a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof.  55. A therapeutic agent for a viral disease, comprising a compound having SH3 domain binding inhibitory activity or a pharmacologically acceptable salt thereof as an active ingredient.  56. Use of a compound having SH3 domain binding inhibitory activity or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for AIDS.  57. Use of a compound having SH3 domain binding inhibitory activity or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for a viral disease.