HK1048110A - NOVEL INTERLEUKIN-1 AND TUMOR NECROSIS FACTOR-α MODULATORS, SYNTHESES OF SAID MODULATORS AND METHODS OF USING SAID MODULATORS - Google Patents
NOVEL INTERLEUKIN-1 AND TUMOR NECROSIS FACTOR-α MODULATORS, SYNTHESES OF SAID MODULATORS AND METHODS OF USING SAID MODULATORS Download PDFInfo
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Description
Technical Field
The present invention relates to compounds and pharmaceutical compositions, including novel compounds and pharmaceutical compositions thereof, useful for the treatment of various diseases and disorders. The invention also relates to methods of synthesizing natural products and novel structurally related compounds. More particularly, the present invention relates to novel analogs of the compounds, methods of making the compounds, and pharmaceutical compositions thereof for treating, for example, inflammation, cancer, cachexia, cardiovascular disease, diabetes, otitis media, sinusitis, and transplant rejection.
Background
Acanthopanax koreanum Nakai (araliaceae) native to Cheju island-korea has been conventionally used as a medicine for treating, for example, neuralgia, paralysis and lumbago. Various useful components have been isolated from the root bark of the tree, including acanthoic acid, a compound having the chemical structure of formula (I). In addition, some analogues of the compounds of formula (I) have been isolated from the root bark of Acanthopanax koreanum Nakai (araliaceae), such as analogues in which the COOH group is substituted with a carbinol group, a methylacetyl ether, a methyl group, and a methyl ester, respectively. See Kim, y.h. and Chung, b.s., j.nat.pro, 51, 1080-83 (1988). (the literature provides the intrinsic chemical names of these analogs). This document, as well as all other cited patents and printed publications, are incorporated herein by reference in their entirety.
The compounds of formula (I), also known as anticancer acids, are reported to have some pharmacological effects, including, for example, analgesic and anti-inflammatory activity. The compounds of formula (I) also show very low toxicity; when administered to rats, the Minimum Lethal Dose (MLD) is 1000 mg/kg. See Lee, Y.S., "(-) -pimaric-9 (11), 15-diene-19-oic acid, a pharmacological study of the composition of Acanthopanax koreanum Nakai," Doctorate Thesis, Dept.of Pharmacy, Seoul National University, Korea (1990). Compounds of formula (I) and/or their natural analogs may exhibit these known pharmacological effects by inhibiting leukocyte migration and prostaglandin E2(PGE2) synthesis, and are suspected to be effectors of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) production. In addition, the preparation of and use of anticancer acids in the treatment of immune disorders is described in International patent publication WO95/34300 (12/21 of 1995).
The compound of formula (IA), kauranoic acid (kauranoic acid), and corresponding methyl ester analogs of the compound of formula (IA), and methanol-reducing analogs of the compound of formula (IA) have been isolated from the root bark of Acanthopanax koreanum Nakai (araliaceae). See alsoKim, y.h. and Chung, b.s., j.nat.pro, 51, 1080 (1988). (this document provides the inherent chemical names of isokaemic acid, (-) -isokauri-16-en-19-oic acid and known analogues of isokaemic acid).
Tumor necrosis factor-alpha (referred to herein as "TNF-alpha" or "TNF") and/or interleukin-1 (referred to herein as "IL-1") are involved in various biochemical pathways, and therefore modulators of TNF-alpha and/or IL-1 activity or production, particularly novel modulators of TNF-alpha and/or IL-1 activity or novel compounds that affect IL-1 or TNF-alpha or IL-1 and TNF-alpha production, are highly desirable. Such a compound or class of compounds would be of value in: maintenance of the human immune system, treatment of diseases such as tuberculous pleurisy, rheumatoid pleurisy, and diseases not generally considered to be immune disorders such as cancer, cardiovascular disease, redness of the skin, viral infections, diabetes, and transplant rejection.
While various methods of modulating the production of tumor necrosis factor-alpha and interleukins are known, new methods, compounds, and pharmaceutical formulations for modulating tumor necrosis factor-alpha and interleukins are highly desirable and have long been sought by those skilled in the art.
Summary of The Invention
It is therefore an object of the present invention to provide synthetic and semi-synthetic processes for the preparation of compounds of formula (I) and (IA) and structural analogs thereof, including novel analogs of the compounds of formula (I) and (IA).
The compounds of the present invention include, for example, compounds having the chemical structure of formula (II) and compounds having the chemical structure of formula (IIA). For compounds having the chemical structure of formula (II), the invention includes:wherein the R groups are defined as follows: if any R3-R5、R7、R8、R11-R13Not being hydrogen, R2Or R6Or R9Not being methyl, or R10Is not CH2Then R is1Selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12An aryl group; however, if all R' s3-R5、R7、R8、R11-R13Are all hydrogen, R2、R6And R9Are each methyl, and R10Is CH2Then R is1Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohols, (C) other than methylacetyl ether1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12And (4) an aryl group.
R2And R9Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Acyl radical, C1-C12Alcohol, and C5-C12And (4) an aryl group.
R3-R5、R7、R8And R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12And (4) an aryl group. In a particularly preferred embodiment, R11Is C1-C6Alkyl, or C1-C6Substituted alkyl, and all other R groups are hydrogen.
R6Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12Alkynyl.
R10Selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group.
R14And R15Each independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6And (4) an aryl group.
For compounds having the chemical structure of formula (IIA), the invention includes:wherein if any R is3-R5、R7、R8、R11-R13Not being hydrogen, R2Or R6Not being methyl, R10Is not CH2Or ifR10Is not CH2OH and R11Not OH, then R1Selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12A substituted alkenyl group; but instead of the other end of the tube
If all R are3-R5、R7、R8、R11-R13Are all hydrogen, R2And R6Are each methyl, and R10Is CH2Or CH2OH, then R1Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohol, (C)1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C2-C12Substituted alkyl, C2-C12Alkenyl, and C2-C12A substituted alkenyl group;
R2selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Acyl radical, C1-C12Alcohol, and C5-C12An aryl group;
R3、R4、R5、R7、R8and R11-R13Are respectively provided withIndependently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12And (4) an aryl group. In a particularly preferred embodiment, R11Is C1-C6Alkyl, or C1-C6Substituted alkyl, and all other R groups are hydrogen.
R6Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12Alkynyl.
R10Selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12An aryl group; and is
R14And R15Can be stereospecific and independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6And (4) an aryl group.
It is another object of the present invention to provide compounds having the chemical structure of formula (IIB), and synthetic and semi-synthetic methods for preparing compounds having the chemical structure of formula (IIB). For compounds having the chemical structure of formula (IIB), such as those referred to herein as TTL1, TTL2, TTL3, TTL4, and analogs and derivatives thereof, the present invention includes:wherein the R groups are defined as follows: r1Selected from hydrogen, halogen, COOH, C1-C12A carboxylic acid,C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group. Under these conditions, R1Preferably selected from COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl, and C1-C12Ester, most preferably selected from COOH and C1-C6And (3) an ester.
R2And R9Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Acyl radical, C1-C12Alcohol, and C5-C12An aryl group;
R3-R5、R7、R8and R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12And (4) an aryl group. In a particularly preferred embodiment, R11Is C1-C6Alkyl, or C1-C6Substituted alkyl, and all other R groups are hydrogen.
R6Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12Alkynyl.
R10Selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group.
R14And R15Are stereospecific and are each independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6And (4) an aryl group.
It will also be appreciated that each R group, particularly R, may be selected3、R4、R5、R7、R8And R11-R13To form a ring system. For example, R13And R12May be ethylene and may include a covalent C-C bond between their respective terminal carbons to form another 6-membered ring in formula (IIB). As a further example, by selecting each R group of formula (IIB), in particular R3、R4、R5、R7、R8And R11-R13May form a bicyclic ring.
The compounds of the present invention include prodrug esters of the compounds of formulas (II), (IIA), and (IIB), and acid addition salts of the compounds of formulas (II), (IIA), and (IIB), as well as pharmaceutical compositions comprising a therapeutically effective amount of the compounds, including prodrug esters and acid addition salts thereof, and optionally a pharmaceutically acceptable carrier. Such compositions are useful, for example, as anti-inflammatory analgesics, for the treatment of immune and autoimmune diseases, as anti-cancer or anti-tumor agents, and for the treatment of cardiovascular disease, skin redness, viral infections, diabetes, otitis media, sinusitis, and/or transplant rejection. In particular, pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (II), (IIA), or (IIB), or a prodrug ester and acid addition salt of a compound of formula (II), (IIA), or (IIB), are useful as anti-cancer agents, anti-tumor agents, anti-viral agents, and for the treatment of cardiovascular disease, skin redness, viral infection, diabetes, otitis media, sinusitis, and/or transplant rejection.
The present invention also provides a novel method for synthesizing the above compound and analogs thereof, comprising carrying out Diels-Alder reaction with a diene having two or more rings and a dienophile compound to produce a compound having 3 or more rings; and generating the desired synthetic compound. The Diels-Alder reaction, and the selectivity of dienes to dienophiles, provides flexibility in the synthesis of various compounds of the invention and enables the use of combinatorial chemical libraries of compounds of the invention for bioassays, including clinical trials.
Brief Description of Drawings
Some preferred embodiments of the invention are illustrated in the accompanying drawings. The drawings illustrate only some preferred embodiments of the invention and/or some preferred methods of making and/or using the invention. The drawings are not intended to limit the scope of the invention described and claimed herein.
FIG. 1 shows the structures of anticancer acid and its methyl ester, the stereochemical view of the anticancer acid, and the skeletal type view of some of the compounds of the present invention.
FIG. 2 shows retrosynthetic analysis and strategic linkage of some of the compounds of the invention.
FIG. 3 shows an alternative method for constructing the AB ring of some of the compounds of the present invention, which comprises: a Wenkert's method for synthesizing (+/-) arhanedioic acid; welch's method for synthesizing (+/-) podocaronic acid; and a DeGrot's method for synthesizing (+/-) arhanedioic acid.
FIG. 4 shows a schematic synthesis scheme for the synthesis of the acontaic acid and the AB ring of some of the compounds of the invention (scheme 1).
FIG. 5 shows a synthetic scheme for carrying out the synthesis of the acetylac acid and some of the compounds of the invention (scheme 2).
FIG. 6 shows a minimal three-dimensional model of a diene 42 as described in the detailed description of the invention.
FIG. 7 shows a synthesis scheme (FIG. 3) for developing and using a catalyst 49 for an asymmetric Diels-Alder reaction as described in detail in the description of the preferred embodiment of the invention.
FIG. 8 shows a synthetic scheme based on the asymmetric Diels-Alder method for the synthesis of compounds of formula (I) and some of the compounds of the invention (scheme 4).
FIG. 9 shows the structure-activity relationship and the focus of the structure-activity relationship research of caryophyllin and derivatives thereof and some compounds of the present invention.
FIG. 10 shows the identified sites for structural changes and structure-activity relationships of Compound 1.
FIG. 11 shows preferred representative examples of analogs of Compound 1 for structure-activity relationship studies and chemo-biological studies.
FIG. 12 shows some preferred derivatives of representative Compound 1 for use in photoaffinity labeling studies.
FIG. 13 shows some preferred representative examples of dimers and/or conjugates of Compound 1.
Figure 14 shows the total chemical synthesis of some of the compounds of the invention identified in figure 17 as TTL1 and TTL 3.
Figure 15 shows the chemical synthesis of the preferred 14C-labelled compounds of the present invention identified as TTL3 in figure 17.
FIG. 16 shows the total chemical synthesis of the compound of formula (I).
Figure 17 is a summary of the synthesis of some of the compounds of the present invention and the physical properties of these compounds. The compounds TTL1, TT12, TTL3, and TTL4 are as defined in the figure.
FIG. 18 shows a summary of the synthesis of example 1.
FIG. 19 shows the structures of (-) antathoic acid and (+) pimaric acid.
FIG. 20 shows the inverse synthetic analysis of (-) antathoic acid of example 1.
FIG. 21 shows a scheme for the synthesis of the preferred compounds of formula (IIB) (scheme 5) as described in examples 1-6. The reagents, conditions and percentage yield of each step were as follows: (a)0.1 equivalent of PTSA (CH)2OH)2Benzene, 80 ℃,4 hours, 90%; (b)2.2 equivalents of Li, liquid NH31.0 equivalent of tBuOH, -78- -30 ℃ for 30 minutes, followed by isoprene (excess), -78-50 ℃; 1.1 equivalent of NC-CO2Me，Et2O, 55 percent at the temperature of-78-0 ℃ for 2 hours; (c)1.1 equivalents of NaH, HMPA, 25 ℃, 3 hours; 1.1 equivalent of MoMCl, 25 ℃,2 hours, 95%; (d)7.0 equivalents of Li, liquid NH320 minutes at-78- -30 ℃; CH (CH)3I (excess), 78- -30 ℃ for 1 hour, 61%; (e)1N HCl, THF, 25 ℃, 15 minutes, 95%; (f)1.6 equivalents of Li acetylide, Et2O, 25 ℃,1 hour, 91%; (g) lindele catalyst (20% by weight), H2Dioxane/pyridine 10/1, 25 ℃, 10 min, 95%; (h)4.4 equivalents BF3·Et2O, benzene/THF 4/1, 80 ℃,5 hours, 95%; (i)13 equivalents of compound 103, pure, 8 hours, 25 ℃, 100%; (j)1.4 equivalents of NaBH4THFMeOH: 10/1, 30 minutes, 25 ℃, 94%; (k)1.1 equivalent of p-Br-C6H4COCl, 1.5 equivalents of pyridine, 0.1 equivalents of DMAP, CH2Cl225 ℃ for 2 hours, 95% for compound 116 and 97% for compound 117.
FIG. 22 shows the Chem3D representation of the ORTEP diagram of compounds 116 and 117, where only selected hydrogen atoms are shown for clarity.
FIG. 23 shows a scheme for synthesizing a tricyclic core of (-) antathoic acid of example 1 (FIG. 6). The reagents, conditions and percentage yield of each step were as follows: (a)3.0 equivalents PhSH, 0.05 equivalents AIBN, xylene, 120 ℃,18 hours, 86%, (b)1.1 equivalents POCl3HMPA, 25 ℃,1 h; 1.1 equivalent of pyridine, 150 ℃,18 hours, 81%; (c)3.0 equivalents of Compound 103, 0.2 equivalents of SnCl4(1M solution in methylene chloride), CH2Cl284% at-20-0 ℃ for 20 hours; (d)1.4 equivalents of NaBH4EtOH, 25 ℃, 30 minutes; (e) raney nickel (excess), THF, 65 ℃, 10 min, 91% (two step yield); (f)1.3 equivalents Dess-Martin periodinane, CH2Cl230 minutes at 25 ℃; (g)2.7 equivalents of P3PhCH3Br, 2.2 equivalents NaHMDS (1.0 in THF), THF, 25 ℃,18 hours, 86% (two step yield); (h)3.0 LiBr, DMF, 160 ℃, 3 hours, 93%.
Detailed description of the preferred embodiments
Some of the compounds of the present invention have the chemical structure shown in formula (II).
The R group in the compound of formula (II) may be selected in the following manner. If (1) any R3-R5、R7、R8、R11-R13Is not hydrogen, (2) R2、R6Or R9Is not methyl, or (3) R10Is not CH2Then R is1Selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group. Under these conditions, R1Preferably selected from COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl, and C1-C12Ester, most preferably selected from COOH and C1-C6And (3) an ester.
However, if (1) all R3-R5、R7、R8、R11-R13Are both hydrogen, (2) R2、R6And R9Are each methyl, and (3) R10Is CH2Then R is1Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohols, (C) other than methylacetyl ether1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12And (4) an aryl group. Under these conditions, R1Preferably selected from C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl, and C2-C12Ester, most preferably selected from COOH and C4-C8And (3) an ester.
R2And R9Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Acyl radical, C1-C12Alcohol, and C5-C12And (4) an aryl group. Preferably, R2And R9Each independently selected from alkyl and alkenyl. Most preferably, R2And R9Each is methyl, but in a preferred embodiment of the compounds of formula (II), R is2And R9One of which may be methyl and the other not.
R3、R4、R5、R7、R8And R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12And (4) an aryl group. Preferably, R3、R4、R5、R7、R8And R11-R13Are each hydrogen or C1-C6Alkyl, most preferably, R3、R4、R5、R7、R8And R11-R13Respectively hydrogen. However, in preferred embodiments of the compounds of formula (II), any one or more of R3、R4、R5、R7、R8And R11-R13May be hydrogen and the others may not be.
R6Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12Alkynyl. R6Preferably selected from hydrogen, halogen, C1-C6An alkyl group. R6More preferably C1-C6Alkyl radical, R6Most preferred is methyl.
R10Selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6SubstitutionAlkenyl of, C1-C12Alcohol, and C5-C12And (4) an aryl group. Connection R10The bond to the remainder of the compound of formula (II) is preferably a C-C double bond, but may be a C-C single bond, a C-H single bond, or a heteroatom single bond. R10Preferably CH2Or CH2R ', wherein R' is C1-C6Alkyl, or C1-C6A substituted alkyl group. R10Most preferably CH2。
R14And R10Each independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6Aryl, wherein hydrogen and C1-C6Alkyl radical, C1-C6Substituted alkyl groups are most preferred.
It will also be appreciated that each R group, particularly R, may be selected3、R4、R5、R7、R8And R11-R13To form a ring system. For example, R13And R12May be ethylene and may include a covalent C-C bond between their respective terminal carbons to form another 6-membered ring in the compound of formula (II). As a further example, by selecting each R group of formula (IIB), in particular R3、R4、R5、R7、R8And R11-R13May form a bicyclic ring.
Some preferred compounds of the invention have the chemical structure shown in formula (IIA).
The R group in the compound of formula (IIA) may be selected in the following manner. If any R3-R5、R7、R8、R11-R13Not being hydrogen, R2Or R6Not methyl, or if R10Is not CH2OH and R11Is OH, then R1Selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12A substituted alkenyl group. Under these conditions, R1Preferably selected from COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl, and C1-C12Ester, most preferably selected from COOH and C1-C6And (3) an ester.
If all R are3-R5、R7、R8、R11-R13Are all hydrogen, R2And R6Are each methyl, and R10Is CH2Or CH2OH, then R1Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohol, (C)1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C2-C12Substituted alkyl, C2-C12Alkenyl, and C2-C12A substituted alkenyl group. Under these conditions, R1Preferably selected from C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl, and C2-C12Esters, most preferably C4-C8And (3) an ester.
R2Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Acyl radical, C1-C12Alcohol, and C5-C12And (4) an aryl group. Preferably, R2And R9Each independently selected from alkyl and alkenyl. Most preferably, R2And R9Each methyl, although in a preferred embodiment of the compound of formula (IIA), R is2And R9One of which may be methyl and the other not.
R3、R4、R5、R7、R8And R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12And (4) an aryl group. Preferably, R3、R4、R5、R7、R8And R11-R13Are each hydrogen or C1-C6Alkyl, most preferably, R3、R4、R5、R7、R8And R11-R13Respectively hydrogen. However, in preferred embodiments of the compounds of formula (IIA), any one or more of R3、R4、R5、R7、R8And R11-R13May be hydrogen and the others may not be.
R6Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12Alkynyl. R6Preferably selected from hydrogen, halogen, C1-C6An alkyl group. R6More preferably C1-C6Alkyl radical, R6Most preferred is methyl。
R10Selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group. Connection R10The bond to the remainder of the compound of formula (IIA) is preferably a C-C double bond, but may be a C-C single bond, a C-H single bond, or a heteroatom single bond. R10Preferably CH2Or CH2R ', wherein R' is C1-C6Alkyl, or C1-C6A substituted alkyl group. R10Most preferably CH2。
It will also be appreciated that each R group, particularly R, may be selected3、R4、R5、R7、R8And R11-R13To form a ring system. For example, R13And R12May be ethylene and may include a covalent C-C bond between their respective terminal carbons to form another 6-membered ring in the compound of formula (IIA). As a further example, by selecting the individual R groups of formula (IIA), especially R3、R4、R5、R7、R8And R11-R13May form a bicyclic ring.
Some preferred compounds of the invention, including compounds referred to herein as TTL1, TTL2, TTL3, and TTL4, have the chemical structure shown in formula (IIB).
The R group in the compound of formula (IIB) may be selected in the following manner: r1Selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group. Under these conditions, R1Preferably selected from COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl, and C1-C12Ester, most preferably selected from COOH and C1-C6And (3) an ester.
R2And R9Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Acyl radical, C1-C12Alcohol, and C5-C12And (4) an aryl group. Preferably, R2And R9Each independently selected from alkyl and alkenyl. Most preferably, R2And R9Each methyl, although in a preferred embodiment of the compounds of formula (IIB), R is2And R9One of which may be methyl and the other not.
R3、R4、R5、R7、R8And R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12And (4) an aryl group. Preferably, R3、R4、R5、R7、R8And R11-R13Are each hydrogen or C1-C6Alkyl, most preferably, R3、R4、R5、R7、R8And R11-R13Respectively hydrogen. However, in preferred embodiments of the compounds of formula (IIB)In (1), any one or several R3、R4、R5、R7、R8And R11-R13May be hydrogen and the others may not be.
R6Selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12Alkynyl. R6Preferably selected from hydrogen, halogen, C1-C6An alkyl group. R6More preferably C1-C6Alkyl radical, R6Most preferred is methyl.
R10Selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group. Connection R10The bond to the remainder of the compound of formula (IIB) is preferably a C-C double bond, but may be a C-C single bond, a C-H single bond, or a heteroatom single bond. R10Preferably CH2Or CH2R ', wherein R' is C1-C6Alkyl, or C1-C6A substituted alkyl group. R10Most preferably CH2。
It will also be appreciated that each R group, particularly R, may be selected3、R4、R5、R7、R8And R11-R13To form a ring system. For example, R13And R12May be ethylene and may include a covalent C-C bond between their respective terminal carbons to form another 6-membered ring in the compound of formula (IIB). As a further example, by selecting each R group of formula (IIB), in particular R3、R4、R5、R7、R8And R11-R13May form a bicyclic ring. Definition of
As used hereinThe term "alkyl" refers to any unbranched or branched saturated hydrocarbon wherein C1-C6Unbranched saturated unsubstituted hydrocarbons are preferred, and methyl, ethyl, isobutyl, and tert-butyl are most preferred. Among the substituted saturated hydrocarbons, C1-C6Mono-and di-and perhalo-substituted saturated hydrocarbons and amino-substituted hydrocarbons are preferred, with perfluoromethyl, perchloromethyl, perfluoro-tert-butyl, and perchloro-tert-butyl being most preferred. The term "substituted alkyl" refers to any unbranched or branched substituted saturated hydrocarbon wherein unbranched C1-C6Secondary alkyl amines, substituted C1-C6Secondary alkylamine, and unbranched C1-C6Tertiary alkyl amines are within the definition of "substituted alkyl" but are not preferred. The term "substituted alkyl" refers to any unbranched or branched substituted saturated hydrocarbon. The cyclic compound, whether a cyclic hydrocarbon or a cyclic compound having a heteroatom, is within the meaning of "alkyl".
The term "substituted" as used herein means any substitution of a hydrogen atom with a functional group.
The term "functional group" as used herein has its general definition and refers to a chemical moiety preferably selected from the group consisting of: halogen atom, C1-C20Alkyl, substituted C1-C20Alkyl, perhaloalkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, benzyl, heteroaryl, substituted heteroaryl, cyano, and nitro. The functional groups may also be selected from: -SRs、-ORo、-NRn1Rn2、-N+Rn1Rn2Rn3、-N＝N-Rn1、-P+Rn1Rn2Rn3、-CORc、C(＝NORo)Rc、-CSRc、-OCORc、-OCONRn1Rn2、-OCO2Rc、-CONRn1Rn2、-C(＝N)Rn1Rn2、-CO2Ro、-SO2NRn1Rn2、-SO3Ro、-SO2Ro、-PO(ORo)2、-NRn1CSNRn2Rn3. Substituents R of these functional groupsn1、Rn2、Rn3、RoAnd RsPreferably independently selected from hydrogen atom, C1-C20Alkyl, substituted C1-C20Alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, benzyl, heteroaryl, substituted heteroaryl, and may constitute part of an aliphatic or aromatic heterocycle. RcPreferably selected from hydrogen atom, C1-C20Alkyl, substituted C1-C20Alkyl, perhaloalkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, benzyl, heteroaryl, substituted heteroaryl, and cyano.
The terms "halogen" and "halogen atom" as used herein refer to any one of the radiation-stable atoms in column 17 of the periodic Table of the elements, preferably fluorine, chlorine, bromine or iodine, with fluorine and chlorine being particularly preferred.
The term "alkenyl" as used herein refers to any unbranched or branched substituted or unsubstituted unsaturated hydrocarbon wherein C is1-C6Unbranched monounsaturated and diunsaturated unsubstituted hydrocarbons are preferred, and monounsaturated dihalosubstituted hydrocarbons are most preferred. The term "substituted alkenyl" refers to any unbranched or branched, substituted unsaturated hydrocarbon substituted with one or more functional groups, wherein unbranched C2-C6Secondary alkenylamines, substituted C2-C6Secondary alkenylamine, and unbranched C2-C6The tertiary alkenyl amines are within the definition of "substituted alkyl". The term "substituted alkenyl" refers to any unbranched or branched substituted unsaturated hydrocarbon. The cyclic compound, whether an unsaturated cyclic hydrocarbon or a cyclic compound having a heteroatom, is within the meaning of "alkenyl".
As used herein, the term "alcohol" refers to any unbranched or branched saturated or unsaturated alcohol, wherein C1-C6Unbranched saturated unsubstituted alcohols are preferred, and methanol, ethanol, isobutanolAlcohols, and t-butanol are most preferred. Among the substituted saturated alcohols, C1-C6Mono-and di-substituted saturated alcohols are preferred. The term "alcohol" includes substituted alkyl alcohols, and substituted alkenyl alcohols.
The term "aryl" as used herein includes the terms "substituted aryl", "heteroaryl", and "substituted heteroaryl", and refers to an aromatic hydrocarbon ring preferably having 5 or 6 ring atoms. The terms "heteroaryl" and "substituted heteroaryl" refer to an aromatic hydrocarbon ring having at least one heteroatom such as oxygen, sulfur, or nitrogen atom and at least one carbon atom in the ring. "aryl" means, most generally, and "substituted aryl", "heteroaryl" and "substituted heteroaryl" more particularly, an aromatic hydrocarbon ring preferably having 5 or 6 ring atoms, most preferably having 6 ring atoms. The term "substituted aryl" includes mono-or poly-substituted aryl groups substituted with, for example, alkyl, aryl, alkoxy, azide, amine, and amino groups. "heteroaryl" and "substituted heteroaryl", if used independently, specifically refer to an aromatic hydrocarbon ring having at least one heteroatom such as oxygen, sulfur, or nitrogen atom and at least one carbon atom in the ring.
The terms "ether" and "alkoxy" refer to any unbranched or branched, substituted or unsubstituted, saturated or unsaturated ether wherein C is1-C6Unbranched saturated unsubstituted ethers are preferred, with dimethyl ether, diethyl ether, methyl isobutyl ether, and methyl tert-butyl ether being most preferred. The terms "ether" and "alkoxy" refer most generally, and "cycloalkoxy" and "cyclic ether" more particularly to non-aromatic hydrocarbons preferably having 5 to 12 ring atoms.
The term "ester" refers to any unbranched or branched, substituted or unsubstituted, saturated or unsaturated ester wherein C is1-C6Unbranched saturated unsubstituted esters are preferred, methyl and isobutyl esters being most preferred.
The term "prodrug ester", especially when referring to a prodrug ester of a compound of formula (I), refers to a derivative of a compound that is rapidly converted in vivo to the original compound, for example by hydrolysis in blood. The term "prodrug ester" refers to a derivative of a compound of the invention formed by the addition of any of several ester-forming groups that are hydrolyzable under physiological conditions. Examples of prodrug ester groups include pivaloyloxymethyl, acetoxymethyl, 2-benzo [ c ] furanonyl, indanyl, and methoxymethyl, as well as other such groups known in the art, including (5-R-2-oxo-1, 3-dioxol-4-yl) methyl. Further examples of prodrug ester groups can be found in, for example, T.Higuchi and V.Stella "Pro-drugs as Novel Delivery Systems", Vol.14, A.C.S.Symposium series, American Chemical Society (1975); and "Bioreversible Carriers in drug design: the Theory and Application ", e.b. roche editors, Pergamon Press: new York, 14-21(1987) (examples of esters useful as prodrugs of carboxyl-containing compounds are provided).
The term "pharmaceutically acceptable salt", especially when referring to a pharmaceutically acceptable salt of a compound of formula (I), refers to any pharmaceutically acceptable salt of a compound, preferably to an acid addition salt of a compound. Preferred examples of pharmaceutically acceptable salts are alkali metal salts (sodium or potassium), alkaline earth metal salts (calcium or magnesium), or salts derived from ammonia or pharmaceutically acceptable organic amines such as C1-C7Ammonium salts of alkylamines, cyclohexylamine, triethanolamine, ethylenediamine or tris (hydroxymethyl) aminomethane. For compounds of the invention which are basic amines, preferred examples of pharmaceutically acceptable salts are acid addition salts of pharmaceutically acceptable inorganic or organic acids, such as hydrohalic acids, sulfuric acid, phosphoric acid, or aliphatic or aromatic carboxylic or sulfonic acids, such as acetic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinic acid, methanesulfonic acid, p-toluenesulfonic acid, or naphthalenesulfonic acid. Preferred pharmaceutical compositions of the present invention comprise pharmaceutically acceptable salts and prodrug esters of the compounds of formulae (II), (IIA), and (IIB).
The terms "pure", "substantially pure", and "isolated" as used herein mean that the compounds of the present invention are free of other, different compounds to which the compounds of the present invention are normally bound in their natural state, such that the compounds of the present invention make up at least 0.5%, 1%, 5%, 10%, or 20%, and most preferably at least 50% or 75% of a given sample weight. In a preferred embodiment, these terms mean that the compound of the invention makes up at least 95% of the weight of a given sample.
The terms "anti-cancer", "anti-tumor" and "tumor growth inhibition" when modifying the term "compound" and/or the term "tumor", and the terms "inhibition" and "attenuation" when modifying the term "compound" and/or the term "tumor", mean that the presence of a compound of the invention results in at least a reduction in the growth rate of the tumor or cancer entity. More preferably, the terms "anti-cancer", "anti-tumor", "tumor growth inhibition", "inhibition" and "attenuation" refer to the presence of a compound of the invention resulting in at least a temporary cessation of tumor growth or growth of a cancer entity. The terms "anti-cancer", "anti-tumor", "tumor growth inhibition", "inhibition" and "attenuation" also mean, in particular in the most preferred embodiment of the invention, that the presence of a compound of the invention results in at least a temporary reduction of tumor mass. These terms refer to cancers and various malignancies in animals, particularly mammals, and most particularly humans.
The term "skin redness" refers to any skin redness, especially chronic skin redness of neurogenic origin, which is consistent with, but not limited to, the meaning given in EP 7744250, which is incorporated herein by reference in its entirety.
The term "viral infection" refers to infection by any virus of origin, including rhinoviruses, and preferably, but not limited to, Human Immunodeficiency Virus (HIV), human cytomegalovirus, hepatitis a virus, hepatitis b virus, and hepatitis c virus.
The term "cardiovascular disease" refers to various diseases of the heart and vascular system including, but not limited to, congestive heart failure, cardiac dysfunction, reperfusion injury, and various known abnormalities of peripheral circulation. By "cardiovascular disease" is meant such a disease in an animal, particularly a mammal, most particularly a human.
The term "diabetes" as used herein refers to a variety of diseases involving high insulin levels, insulin resistance, or diabetes, including type I diabetes, type II diabetes, and various related conditions, including but not limited to Stein-Leventhal syndrome and polycystic ovary syndrome (PCOS).
The term "transplant rejection" as used herein refers to the conditions and associated symptoms referred to as allograft rejection, xenograft rejection, and autograft rejection, and in preferred embodiments of the invention, refers to human-human allograft rejection.
The term "modulator" or "modulation" as used herein refers to the ability of a compound or therapeutic process to alter the presence or production of a modulated agent, particularly TNF- α or IL-1, in a subject. Most preferably, "modulator" or "modulation" refers to the ability of a compound or therapeutic process to reduce the presence or production of a modulated species.
As used herein, the terms TTL1, TTL2, TTL3, TTL4 and TTL5 refer to the particular chemical entities identified in figure 17, among other figures.
All other chemical, medical, pharmacological, or technical terms used herein should be understood as understood by one of ordinary skill in the art. Interleukin-1 (IL-1) is a regulatory factor involved in a variety of mammalian immune and inflammatory mechanisms as well as other defense mechanisms, particularly in humans. See, e.g., Dinarello, d.a., FASEB j., 2, 108 (1988). IL-1, the earliest discovered as a factor produced by activated macrophages, is secreted by a variety of cells such as fibroblasts, keratinocytes, T cells, B cells, and brain astrocytes, and is reported to have a variety of functions including: stimulation of CD4+T cell proliferation, see Mizel, s.b., immunol.rev., 63, 51 (1982); stimulation of thymic TcCell killing of cells via their binding to the T cell receptor TCR, see McConkey, d.j., et al, j.biol.chem., 265, 3009 (1990); induction of various substances involved in inflammatory mechanisms such as PGE2Phospholipase A2(PLA2) And collagenase production, see Dejana,e, et al, Bolid, 69, 695-699 (1987)); induction of acute phase protein growth in the liver, see Andus, t., et al, eur.j.immunol., 123, 2928 (1988)); increasing blood pressure in the vascular system, see Okusawa, s., et al, j.clin.invest., 81, 1162 (1988)); and induce the production of other cytokines such as IL-6 and TNF- α, see Dinarello, c.a., et al, j.immunol., 139, 1902 (1987). IL-1 modulation is also known to act on rheumatoid arthritis, see Nouri, a.m., et al, clin.exp.immunol, 58, 402 (1984); transplant rejection, see Mauri and Teppo, Transplantation, 45, 143 (1988); and sepsis, see Cannon, j.g., et al, lymphkin res., 7, 457(1988), and IL-1 can cause fever and pain when administered in large doses, see Smith, j., et al, am.
The incidence of sepsis, arthritis, inflammation and related conditions in animal models can be reduced by inhibiting IL-1 binding to its receptor using natural IL-1 receptor inhibitors (IL-1 Ra), see Dinarello, c.a. and Thompson, r.c., immunol.today, 12, 404(1991), and several methods have been proposed to inhibit IL-1 activity using specific antibodies, see Giovine, d.f.s. and Duff, g.w., immunol.today.11, 13 (1990). For IL-6, proliferation of myeloid cells in myeloma patients caused by excessive secretion of IL-6 has been inhibited by using antibodies against IL-6 or IL-6 receptor, see Suzuki, H.J., Immuno, 22, 1989 (1992). Diseases that can be treated according to the present invention by modulating TNF- α and IL-1 using the compounds of the present invention include, but are not limited to, the diseases described herein. Tumor necrosis factor-alpha (TNF-alpha)
Human TNF-. alpha.was first purified in 1985. See Aggarwal, b.b.; kohr, w.j. "human tumor necrosis factor, production, purification and characterization", j.biol.chem.1985, 260, 2345-2354. Shortly thereafter, molecular cloning of the TNF cDNA and cloning of the human TNF locus were completed. See Pennica, d.; nedwin, g.e.; hayflick, j.s. et al, "human necrosis factor: precursor structure, expression and homology to lymphotoxin ", Nature 1984, 312, 724-729. Wang, a.m.; creasy, A.A.; ladner, m.b. "molecular cloning of complementary DNA of human tumor necrosis factor". Nature1985, 313, 803-806. TNF- α is a trimeric 17-KDa polypeptide, produced primarily by macrophages. The peptide is initially expressed as a 26-kDa transmembrane protein from which the 17-kDa subunit is cleaved off and released after proteolytic cleavage by an enzyme known as TACE. This work revealed a very broad and diverse biological event in which TNF- α was involved and promoted the development of therapies targeting overproduction.
Tumor necrosis factor-alpha (TNF- α) is typically produced by a variety of cells, such as activated macrophages and fibroblasts. TNF- α has been reported to induce IL-1 production, see Dinarello, d.a., fasebj, 2, 108(1988), killing fibrosarcoma L929 cells, see Espevik and Nissen-Meyer, j.immunol. methods, 95, 99 (1986); stimulation of fibroblast proliferation, see Sugarman, b.j., et al, Science, 230, 943 (1985); induces the production of PGE2 and arachidonic acid, both of which can be involved in inflammatory responses, see sutthys, et al, eur.j.biochem., 195, 465 (19991); and induction of IL-6 or other growth factor production, see Van Hinsbergh, et al, Blood, 72, 1467 (1988)). TNF- α has also been reported to be involved directly or indirectly in various diseases such as infectious diseases caused by trypanosoma strains of the genus plasmodium, see Cerami, a., et al, immunol. today, 9, 28 (1988)); immune diseases such as Systemic Lupus Erythematosus (SLE) and arthritis, see Fiers, w., FEBS, 285, 199 (1991); acquired immunodeficiency syndrome (AIDS), see Mintz, M, et al, am.j.dis.child., 143, 771 (1989); sepsis, see Tracey, k.j., et al, curr.opin.immunol., 1, 454 (1989); and some types of infection, see Balkwill, f.r., Cytokines in cancer Therapy, Oxofrd University Press (1989). TNF-alpha and immune response
Infection and tissue damage cause a cascade of biochemical changes that trigger the immune system to begin a chaotic response, known as an inflammatory response. The progression of this response is based, at least in part, on local vasodilation or increased vascular permeability and activation of the vascular endothelium, which allows white blood cells to efficiently circulate and migrate to the site of injury, thereby increasing their chances of binding and destroying any antigen. It is believed that the vascular endothelium is then activated or inflamed. Inflammation is often a welcome immune response to various accidental stimuli, and manifests itself by a rapid onset and short duration (acute inflammation). However, its sustained or uncontrolled activity (chronic inflammation) has a detrimental effect on the body and leads to the onset of several immune diseases, such as: septic shock, rheumatoid arthritis, inflammatory bowel disease, and congestive heart failure. See "tumor necrosis factor-molecules and their emerging role in medicine" b.beutler, ed., Raven Press, n.y.1992, pages 1-590.
The resolution of an effective immune response generally requires the recruitment of various cells and the coordination of a series of biological events. This complex intercellular coordination and interaction is mediated by a group of locally secreted low molecular weight proteins, commonly referred to as cytokines. These proteins bind to specific receptors on the cell surface and initiate signaling pathways that ultimately alter gene expression in target cells, thereby modulating potent inflammatory responses.
Cytokines can exhibit pleiotropic profiles (a given protein exerts different effects on different cells), redundancy (two or more cytokines mediate similar functions), synergy (the combined effect of two cytokines is greater than the additive effect of each individual protein), and antagonism (the effect of one cytokine inhibits the effect of another cytokine). To this end, some cytokines are pro-inflammatory (induce inflammation) while others are anti-inflammatory (inhibit inflammation). Proinflammatory cytokines include: interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha). See "tumor necrosis factor-molecules and their emerging role in medicine" b.beutler, ed., Raven Press, n.y.1992, pages 1-590. These cytokines are secreted by macrophages immediately after the initiation of the inflammatory response and cause coagulation, increase vascular permeability, and activate the expression of adhesion molecules on vascular endothelial cells (e.g., TNF- α stimulates the expression of E-selectin, which binds to neutrophils and recruits them to the site of injury). Subsequently, and during more systemic immune responses, these cytokines act on several organs of the body, including the bone marrow and liver, to ensure increased production of leukocytes and synthesis of appropriate hormones and acute phase proteins. In addition, they act on the hypothalamus and cause fever, thereby helping to inhibit pathogen growth and enhance the overall immune response. TNF-alpha and the pathogenesis of various diseases and disorders
Like many other cytokines, TNF- α is neither completely beneficial nor completely detrimental to the host. But rather maintains a balance of its production and regulation to ensure that the host can effectively react to invading microorganisms without harming the host during the process. As an inflammatory mediator, TNF- α helps the body against bacterial infections and tissue damage by raising the appropriate immune response. However, its overproduction leads to chronic inflammation, has deleterious effects on the body, and plays a major role in the pathogenesis of several diseases, some of which are summarized below.
The disease typically occurs following infection with some gram-negative bacteria such as E.coli, E.aerogenes, and Diplococcus meningitidis. These bacteria carry on their cell walls some lipopolysaccharides (endotoxins) which stimulate macrophages to overproduce IL-1 and TNF- α, thereby causing septic shock. Symptoms of this disorder are often fatal and include a drop in blood pressure, fever, diarrhea, and extensive blood clotting. In the united states alone, about 500,000 people per year suffer from the condition and cause over 70,000 deaths. The annual cost for treating the disease is estimated to be $5-10 billion.
Rheumatoid arthritis, the most common autoimmune disease in humans, affects about 1% of the western population and is the leading cause of disability, the severe form of which leads to death. See Szekanecz, z.; kosh, a.e.; kunkel, s.l.; striiter, r.m. "cytokines in rheumatoid arthritis. potential target for drug use". Clinical pharmacol.1998, 12, 377-390. Camussi, g.; lupin, e. "role of anti-tumor necrosis factor products in the future treatment of rheumatoid arthritis. The condition is characterized by inflammation of the synovium and cellular proliferation, leading to invasion of the adjacent cartilage matrix, which subsequently erodes, ultimately destroying the bone. Although the origin of this inflammatory response is not yet clear, increased TNF-. alpha.and IL-1 expression has been found around areas of cartilage erosion. Recently, the pathogenic role of TNF- α in this disorder has been well studied and experimentally confirmed. Furthermore, clinical data suggest that counteracting the effects of TNF- α may be a therapeutic approach to reduce this erosive process. However, to date, current treatments, although temporarily relieving the disease, do not alter the underlying mechanisms of the disease process or course.
Such diseases, including crohn's disease and ulcerative colitis, are debilitating diseases characterized by chronic inflammation of the intestinal mucosa and lamina propria (lamina propria). Although the events that trigger their initiation are unknown, they are associated with significant leukocyte infiltration and local generation of soluble mediators. Thus, TNF- α is considered to be a key mediator in the pathogenesis of these disorders, acting either through direct cytotoxic effects or as a coordinator of the inflammatory cascade. See, e.g., Armstrong, a.m.; gardiner, k.r.; kirk, s.j.; m.j. hallidar: rowlands, B.J. "tumor necrosis factor and inflammatory bowel disease". Brit.J. surgery 1997, 84, 1051-1058. Data based on acceptable animal models also support theories regarding therapeutic studies in human IBD aimed at reducing the effects of TNF. See Van Deventer, s.j.h. "tumor necrosis factor and crohn's disease" Gut, 1997, 40, 443.
Activated cytokines, in particular TNF- α, are present in patients with chronic heart failure and acute myocardial infarction. See Ferrari, r. "tumor necrosis factor in CHF: bifacial cytokines ". Cardiovasular Res.1998, 37, 554-559. Furthermore, it was demonstrated that TNF- α induces apoptosis processes in cardiac muscle cells both directly (by binding to and genetically reprogramming these cells) and indirectly (by local NO production which also leads to cell death).
HIV replication is activated by the inducible transcription factor NF-. kappa.B, which in turn is induced by TNF-. alpha.. THF induces HIV expression in macrophage and T-cell lines chronically infected with the virus. In a small number of patients with AIDS-related kaposi's sarcoma, infusion of recombinant TNF appears to cause an increase in the level of HIV p24 antigen, a marker of viral replication activity. See "cytokine therapy modulation" CRC Press, Inc., N.Y.1996, pages 221-236. These results provide a mechanistic basis for considering the use of TNF blockers to reduce the burden of HIV infection.
There is evidence that the number of disorders in which TNF is involved is consistently increasing. "cytokine therapy modulation" CRC Press, Inc., N.Y.1996, pages 221-236. In certain situations, such as transplantation, graft-versus-host disease, and ischemia/reperfusion injury, a possible pathogenesis affects the pro-inflammatory activity of TNF- α on various tissue cells. The reduced insulin response in other, e.g., non-insulin dependent diabetes mellitus, involves a more selective TNF- α action, which appears to be outside the scope of the standard proinflammatory model. TNF- α has been detected locally in patients with otitis media (with or without exudative inner ear infections), see e.g., Willett, d.n., Rezaee, r.p., bill, j.m., Tighe, m.a. and dematia, t.f., an.rhinol laryngol, 107 (1998); maxwell, k., Leonard, g., and Kreutzer, d.l., archorylgorl Head new Surg, vol.123, p.984(sept.1997), and TNF- α has also been detected locally in patients with sinusitis, see, e.g., Nonoyana, t.a., Harada, t.a., shinagi, j., Yoshimura, e.g., Sakakura, y.a., Auris Nasus Larynx, 27(1), 51-58(Jan 2000); buehring i., Friedrich b., Schaff, j., Schmidt h., Ahrens p., Zielen s., CLin expimemul, 109(3), 468-472, Sept 1997). TNF-alpha and IL-1 modulation as a method of treatment
The methods employed to treat the above-mentioned disorders will be aimed at reducing chronic inflammation and are steroid and non-steroid based anti-inflammatory treatments before TNF- α is isolated. However, recent our understanding of TNF- α has led us to develop alternative strategies based on its selective inhibition. These general strategies are summarized below.
Such treatments, including the use of corticosteroids, cause a reduction in the number and activity of cells of the immune system. The mechanism of action of corticosteroids involves traversing the plasma membrane and binding to receptors in the cytoplasm. The resulting complexes are then transported into the nucleus where they bind to specific regulatory DNA sequences and thereby down-regulate cytokine production. Although currently being adopted, this strategy has several drawbacks, since it is not specific for TNF- α, but also down regulates several other cytokines that may play an important role in an effective immune response. In addition, the use of steroids may lead to the formation of cancer (e.g. prostate cancer).
Non-steroidal anti-inflammatory treatments. This strategy involves the use of compounds that indirectly reduce inflammation, such as aspirin. This is generally achieved by inhibiting the cyclooxygenase pathway that produces prostaglandins and thromboxanes. This effect reduces vascular permeability and provides temporary relief. In this regard, this strategy does not regulate cytokine production, and has little or no effect on diseases associated with chronic inflammation.
Engineered monoclonal anti-TNF antibodies. This strategy involves the selection of monoclonal antibodies that bind TNF- α and counteract its effects. Although preliminary clinical trials have shown some positive results, this strategy is still in its infancy and not generally accepted. One problem to be addressed is that monoclonal antibodies are of murine origin and in humans they elicit an anti-immunoglobulin immune response, which limits their use. Recombinant engineering techniques are being carried out to generate "humanized" variants of rodent antibodies that maintain anti-TNF- α activity and are more readily accepted by the human immune system.
The use of soluble receptors against TNF-alpha is a new therapeutic approach. While these receptors have been created to bind TNF- α and counteract its effects, they also increase its activity by prolonging its useful life in the blood circulation. In addition, such treatments are estimated to have a long-term immune response.
The goal of this approach is to reduce inflammation not by reducing TNF- α expression, but by locally increasing the production of anti-inflammatory cytokines. Such treatment involves direct injection of a cDNA expression vector encoding an anti-inflammatory cytokine that antagonizes TNF action into the inflamed area. The efficacy of this approach is currently being evaluated in preclinical trials, and its long-term impact on the immune response is unknown.
Furthermore, it has recently been determined that TNF- α and/or IL-1 are involved in the regulation of angiogenic Vascular Endothelial Growth Factor (VEGF), see E.M. Paleolog et al, Arthritis & rhematosis, 41, 1258(1998), and possibly tuberculous pleurisy, rheumatoid pleurisy, and other immune disorders, see T.S * derblom, Eur. Respir. J., 9, 1652 (1996). TNF- α has also been reported to affect the expression of several cancer cell genes for multidrug resistance-associated proteins (MRP) and Lung Resistance Protein (LRP), see v.stein, j.nat. canc.inst., 89, 807(1997), and involvement in chronic and congestive heart failure, as well as related heart diseases, see e.g., r.ferrari, cardiovasular res, 37, 554 (1998); ceconi et al, prog.cardiovasular dis, 41, 25(1998), and direct or indirect mediation of viral infection, see d.k.biswas, et al, j.acquired Immune Defic syndr.hum retrovirol, 18, 426-34(1998) (HIV-1 replication); r.lenauor, et al, res.virol, 145, 199-207(1994) (same); t.harrer, et al, j.acquir Immune defic.syndr., 6, 865-71(1993) (same); fietz, et al, Transplantation, 58(6), 675-80(1994) (human Cytomegalovirus (CMV) regulation); zhang, et al, chi.med.j., 106, 335-38(1993) (HCV and HBV infection). In addition, antagonists of TNF- α have also been shown to be useful in the treatment of skin redness of neurogenic origin, see European patent EPO-774250-B1(De Lacharriere et al).
It has also been determined that TNF- α levels are elevated in persons diagnosed with obesity or exhibiting insulin resistance and are therefore modulators of diabetes. See hotemisliigil, g., Arner, p., Atkuinson, r., Speigelman, b. (1995), "increased adipose tissue expression of tumor necrosis factor- α (TNF- α) in obese and insulin resistant patients". j.clin.invest.95: 2409-2415. TNF-alpha was also identified as an important modulator of graft rejection. See Imagawa, d., Millis, j., Olthofff, k., Derus, l., Chia, d., Sugich, l., Ozawa, m., Dempsey, R., Iwaki, y., Levy, p., Terasaki, p., Busuttil, R. (1990) "the role of tumor necrosis factor in allograft rejection" Transplantation, vol.50, No.2, 219-225.
These observations highlight the importance and desirability of identifying new strategies and/or new compounds and classes of compounds that can selectively affect TNF- α/IL-1 production. Therefore, small molecules that selectively inhibit these cytokines are of particular medical and biological importance, for example in maintaining an active immune system and treating inflammatory diseases. Preferred synthetic methods of the invention
Some embodiments of the invention include novel methods of preparing compounds having the chemical structure of formula (II), (IIA), or (IIB), and novel methods of preparing known analogs of known compounds having the chemical structure of formula (II), (IIA), or (IIB), such as compounds of formula (I) and (IA).
The compounds of the present invention, particularly compounds having the chemical structure of formula (II), (IIA), or (IIB), can be prepared by synthetic or semi-synthetic means. If prepared synthetically, commonly available starting materials may be used, including but not limited to bicyclic compounds having a reactive halide moiety. The compounds of the present invention having at least three rings can be synthesized according to various ring-closure reactions. Such reactions include, but are not limited to Diels-Alder reactions and Dickman condensation reactions. The Diels-Alder reaction preferably comprises reaction of a diene with a substituted alkenyl moiety, such that a third ring of the desired compound is formed. After the dieckmann condensation reaction, the resulting cyclic ketone may preferably be partially reduced. After such synthetic procedures and other well known procedures have been performed, the compounds of the invention can be purified and isolated using procedures such as chromatography or HPLC as well as procedures well known to those skilled in the art.
Alternatively, in accordance with the present invention, compounds having the chemical structures of formulae (I) and (IA) as well as some specific analogs and derivatives thereof can be extracted and isolated from the root bark of Acanthopanax koreanum Nakai, at least in the form of a crude extract comprising acanthoic acid. Such an extract can be preferably obtained according to the following method:
about 1 kg of dried a. koreanum Nakai root bark was obtained, chipped and covered with 1L-3L, preferably 2L of a suitable solvent, most preferably methanol. The mixture is kept at 20-60 ℃ and may be kept at room temperature for at least 10 hours, preferably 12 hours. The mixture was then filtered to remove and retain the filtrate. This operation is repeated, preferably at least 2 times, and the combined filtrates are concentrated under reduced pressure to obtain an extract.
About 100 grams of the extract is partitioned with 200mL to 400mL, preferably 300mL, of an aqueous solution, preferably water, and 200mL to 400mL, preferably 300mL, of an organic solution, preferably diethyl ether. The organic fraction was separated therefrom and then concentrated under reduced pressure to obtain a further extract. The further extract is purified, preferably by column chromatography, more preferably using a silica gel column, using a mixture of suitable organic solvents, preferably a mixture of hexane and ethyl acetate as eluent, to obtain isolated acanthoic acid.
The isolated compounds of formula (I) and (IA) may then be modified by synthetic means to obtain some of the compounds of the invention, in particular compounds having the chemical structure of formula (II) or (IIA). For example, esters R of anionic acids1The analogs can be formed upon the acid catalyzed nucleophilic addition reaction of an alkyl alcohol with the carboxylic acid moiety of an aconthoic acid. Ether R of acoNToic acid1The analogs can be made from primary alkyl halides or alcohols according to Williamson ether synthesis, or by partial reduction of the primary alkyl group. alkyl of acoNToic acidAlkenyl, and alcohol R10Analogs can be formed by catalytic hydrogenation of the alkenyl group, or by electrophilic addition of preferably HCl or HBr or other suitable alkyl halides. Substituted analogues of the acoNToic acid at other R positions may be formed by substitution reactions involving the use of alkyl halides, provided that appropriate reactive groups and associated protecting groups are employed to facilitate the desired reaction. Given the description of all compounds of the present invention provided herein, the preparation of a full range of compounds of the present invention is within the knowledge of one skilled in the art, in light of these and other well-known synthetic reactions.
Described herein are total synthetic methods for preparing the compounds of the present invention, including compounds of general formulae (I), (IA), (II), (IIA), and (IIB). The synthesis includes one or more retrosynthetic analyses of canthaic acid and analogs thereof, synthesis of radiolabeled canthaic acid and analogs thereof, synthesis of dimers and conjugates of compounds of general formula (I), (IA), (II), (IIA), and (IIB). It will also be appreciated by those skilled in the art that these methods are also fully applicable to the preparation of isokaemic acid and its analogs. Compounds of formula (I) and natural analogs thereof
Root bark of Acanthopanax koreanum Nakai (Araliaceae) native to Cheju island-Korea has been traditionally used as a tonic and sedative agent, and a drug for treating rheumatism and diabetes. During the research of this folk medicine, Chung and collaborators identified two new tricyclic diterpenes from their pharmacologically active extracts: the antanthoic acid (compound 1) and its methyl ester (compound 2) as depicted in figure 1. See Kim, y.h.; chung, b.s.; sankawa, u. "pinadiene diterpenes extracted from Acanthopax Koreanum", j.nat. prod.1988, 51, 1080-1083. AcANTHOIC acid is pimarane (3). However, unlike other members of the pimaranes family, 1 has a distinctive stereochemical relationship between C8 and C10, providing a unique pattern of BC ring system attachment.
Prior to the present invention, there was no total chemical synthesis for the preparation of compounds having the structure of formula (I) or analogs thereof. Importantly, the chemical structure of formula (I), 1, (figure 1) has biological properties as an anti-inflammatory agent. More specifically, in vitro studies with activated (inflamed) monocytes/macrophages showed that treatment with 1 (about 0.1 to about 1.0 μ g/ml for 48 hours) resulted in approximately 90% inhibition of TNF-. alpha.and IL-1 production. The inhibition is concentration-dependent and cytokine-specific, since under the same conditions the production of IL-6 or IFN- γ (interferon- γ) is not affected. The in vivo effect of anticathoic acid was evaluated in mice with silicosis (chronic lung inflammation) and liver cirrhosis (liver inflammation and liver fibrosis). Histological analysis showed that treatment with compound 1 resulted in a substantial reduction of fibrotic rheumatoid granuloma and a significant recovery of the cirrhosis hepatocytes. These dramatic results may be attributed, at least in part, to the 1-mediated inhibition of pro-inflammatory cytokines such as TNF- α and IL-1. Compound 1 also showed very little toxicity in mice and was only administered orally at high concentrations (LD > 300mg/100g body weight). See Kang, h. -s.; kim, y. -h.; lee, c. -s; lee, j. -j.; choi, i.; pyun, K. -H., Cellular Immunol.1996, 170, 212-221. Kang, h. -s.; song, h.k.; lee, j. -j.; pyun, K. -H.; choi, I., Mediators inflam.1998, 7, 257-259.
Thus, the chemical structure of formula (I) has potent anti-inflammatory and anti-fibrotic effects and reduced expression of TNF- α and IL-I. Acanthonic acids can therefore be used as chemical prototypes for the development of the novel compounds of the invention. Retrosynthetic analysis of Compounds of formulae (I), (II) and (IIB)
Compounds of formula (I), (II) and (IIB), and preferably compounds of formula (I) and compounds of formula (IIB) referred to herein as TTL1, TTL2, TTL3 and TTL4, may be synthesized in accordance with the present invention. The bond of the compound of formula (I) is broken as shown in figure 2. The new structural arrangement of the BC ring and the presence of a tetravalent C13 center are unusual structures and lead to a new strategy as an aspect of the present invention. The structure is fixed in one step to the desired stereochemistry by the diels-alder method. Dienes, such as diene 14, and dienophiles, such as 15 (Y: oxazolidinone-based adjuvant) are suitable starting materials for carrying out endo-selective diels-alder reactions. To further ensure the desired regiochemical result of the cycloaddition, the diene 14 is temporarily functionalized with a heteroatom (e.g., X ═ OTBS or SPh), and then the heteroatom is removed from the product 13. The endo-preference of the reaction, which is commonly observed, is used to predict the stereochemical relationship between C12 and C13 as shown in product 13, while the diastereoselectivity of the process can be controlled with chiral auxiliaries at the carbonyl center of the dienophile or using chiral catalysts. See Xiang, a.x.; watson, d.a.; ling.t.; theodorakis, E.A. "Total Synthesis of Clerocidin via a novel enantioselective homoallylboronic acid Process". J.org.chem.1998, 63, 6774-6775.
The diene 14 may be formed by palladium (0) catalyzed construction of a C8-C11 bond, which shows that ketone 16 is a synthetic precursor thereof. This ketone is formed from the known Wieland-Miescher ketone (17), and ketone 17 can be conveniently prepared by condensing methyl vinyl ketone (19) with 2-methyl 1, 3-cyclohexanedione (18) (FIG. 2).
In one aspect of the invention, it is recognized that the functionality and relative stereochemistry of the AB ring system of the aconthoic acid (1) is analogous to the structure of podocaronic acid (20). See "total synthesis of natural products" ApSimon, ed.; john Wiley & Sons, Inc., 1973, Volume 8, pages 1-243. Among several synthesis strategies towards 20, highlighted is the synthesis of our proposed 1 as shown in fig. 5. In accordance with the present invention, these methods enable the prediction of the stereochemical outcome of the synthesis of compounds of formula (I), (II), and the opposite stereochemistry of compounds of formula (IIB) and of compounds of formula (IIB) referred to herein as TTL1, TTL2, TTL3, and TTL 4. Total synthesis of Compounds of formulae (I), (II) and (IIB)
The initial step in the synthesis of the antathoic acid (1) and all compounds of formulae (I), (II) and (IIB) involves the (17) reaction of Wieland-Miesher ketone. This compound can be conveniently prepared from compounds 18 and 19 as separate enantiomers via michael addition/Robinson cyclization using catalytic amounts of (R) -proline. The more basic C9 carbonyl group of 17 is selectively protected and ketene 34 is then reductively alkylated with methyl cyanoformate,to produce the keto ester 36. The conversion of 36 to 39 was based on previous studies, see Welch, s.c., as shown in figure 3; hagan, C.P. "New stereoselective method for the formation of the A ring of a Rohan's pine acid compound" Synthetic Commun.1972, 2, 221-225. Reduction of the ester function of 39, followed by silylation of the resulting alcohol, followed by deprotection of the acid-catalyzed ketal unit, affords ketone 40. Conversion of 40 to the desired diene 42 is carried out in two steps, including conversion of 40 to its corresponding enol triflate derivative, followed by palladium catalyzed coupling with vinyl stannane 41, see Farine, v.; hauck, s.i.; firestone, r.a. "synthesizes cephems" Bioorg bearing an olefinic sulfoxide side chain for use as a potential beta-lactamase inhibitor.& Medicinal Chem.Lett.1996，6，1613-1618。
The steps for completing the synthesis of the enantiomeric acid (1), and for the synthesis of formulae (I), (II) and (IIB) are depicted as synthetic scheme 2 in FIG. 5. Diels-Alder cycloaddition of diene 42 to dienophile 43 is carried out followed by reductive desulfurization with Raney nickel to produce tricyclic system 44 having the desired stereochemistry. Conversion of 44 to the Weinreb amide followed by DIBALH reduction produces aldehyde 45, which aldehyde 45 is subjected to wittig reaction to produce olefin 46. 46 is subjected to fluoride-induced desilylation, and the resulting alcohol is then subjected to a two-step oxidation to produce the carboxylic acid and make the antathoic acid (1) and useful for preparing compounds of formulae (I), (II) and (IIB) by appropriate substitution of the intermediate.
An important step in the synthesis of compounds of formula (I) and (IA), and compounds of formula (II), (IIA) and (IIB) is the Diels-Alder reaction. This reaction, and the use and selection of one or more appropriately substituted dienes and/or dienophiles, enables the selective synthesis of compounds of formula (II) or of compounds of formula (IIB). For example, as described for example in FIGS. 5,7, 8, 21 and 23 as reaction schemes 2, 3, 4, 5 and 6, the preferred dienophiles described below can be used in place of the dienophiles, e.g., compound 43 and pimarane (103), toSelectively producing compounds of formula (II) and (IIB). Examples of dienophiles include dienophiles of formula (III):wherein the numbered R groups (R)9、R14And R15) The unnumbered R group may be any R as defined above for compounds of formula (IIB)1-R15。
In addition, the electronic conformation of dienes such as compound (42) and compound (112) as described, for example, in fig. 5,7, 8, 21 and 23 as reaction schemes 2, 3, 4, 5 and 6, respectively, can be altered by covalently attaching an electron donating or electron withdrawing group such as pHS to the diene. For example, such covalently attached electron donating or electron withdrawing groups affect the orientation of the newly introduced dienophile.
Thus, in accordance with one aspect of the present invention, the chiral nature of the diene 42 makes it useful for inducing asymmetry during cycloaddition. The minimization model of 42 was determined and the results showed that the angle methyl group at C10 affected the surface selectivity of the reaction and a more efficient dienophile process was performed from the upper surface of the diene. The process produces an adduct which results in the formation of a compound of formula (IIB). The method also enables the development of a catalytic asymmetric variant of the Diels-Alder reaction. In contrast to chiral auxiliaries, the effect of using chiral catalysts is evident and is well documented in recent literature.
One preferred embodiment of the present invention is the use of catalyst 49, developed by Corey and used to improve the asymmetric synthesis of cassio (synthesis scheme 3). See Corey, e.j.; imai, n.; zhang, h. -y.j.am.chem.soc.1994, 116, 3611. It was shown that compound 49 allowed Diels-Alder cycloaddition of the electron rich diene 47 with methacrolein (48) to proceed and produced only the endo-adduct in good yield and enantiomeric excess (83% yield, 97% ee).
The use of the above method in our synthesis is depicted in FIG. 8 as scheme 4. The use of catalyst 49 provides additional versatility and greatly reduces the total number of steps required to complete the 1-up synthesis. Synthesis of radiolabeled Compounds of formula (I)
Radiolabeled samples of compounds of formula (I), (II), (IIA) or (IIB) can be synthesized and used for pharmacological and pharmacokinetic testing, for example, by incorporating a C14-labeled methylene carbon onto a compound of formula (I) using aldehyde 52 as the starting material (FIG. 4 ). The C14-labeled material required for the Wittig chemistry can be prepared in two steps from C14-labeled iodomethane and triphenylphosphine and then treated with a base such as NaHMDS. Base-induced deprotection of the methyl ester produces a radiolabelled compound of formula (I), (II), (IIA) or (IIB).Purpose of synthesizing compounds of formula (II), (IIA) and (IIB)
One aspect of the present invention is the identification of novel anti-inflammatory agents having the structure of compounds of formulae (II), (IIA) and (IIB). The biological screening of the synthetic intermediates and the rationally designed compounds of formula (II) provides information and guides design requirements.
The design and synthesis of analogues of the compounds of formula (II) is based on the following objectives: (a) determining the minimum structural and functional requirements (minimum pharmacophore) for compounds of formula (II) that confer TNF- α and IL-1 modulating activity; (b) improving the TNF-alpha and IL-1 modulating activity of the compound of formula (II) by modifying the structure, particularly the R group of the minimal pharmacophore (e.g. SAR studies and molecular recognition experiments); (c) determining the mode of action of the compound of formula (II) by a photoaffinity labeling assay; (d) altering and improving the solubility and membrane permeability of the compound of formula (II); (e) synthesizing and assaying a conjugate of a dimer of a compound of formula (II); selective delivery units, and (f) redesigning and refining the target structure by evaluating the resulting biological data.
Of particular interest in the rational design of compounds of formulae (II), (IIA) and (IIB) is the recent report that modification of the A and C rings of oleanolic acid (53) as shown in FIG. 9 enhances antiproliferative and anti-inflammatory activity. See Honda, t.; rounds, b.v.; grible, g.w.; suh, n.; wang, y.; sporn, M.B. "design and synthesis of 2-cyano-3, 12-dioxaoleanane-1, 9-dien-28-oic acid, a novel highly active inhibitor that inhibits nitric oxide production in murine macrophages" Biorg. & medicinal. chem.Lett.1998, 8, 2711-2714. Suh, N. et al "A novel oleanane triterpene compound 2-cyano-3, 12-dioxaoleanane-1, 9-diene-28-oic acid" Cancer Res.1999, 59, 336-341 having potent differentiation, antiproliferative and anti-inflammatory activities. More specifically, the SAR studies carried out with the commercially available 53 and its semisynthetic derivatives have led us to the realization that: (a) the biological effectiveness of 53 can be increased by attaching an electron withdrawing group such as nitrile to the C2 position (FIG. 9); (b) the alpha, beta unsaturated ketone functionality on the C ring is a strong efficacy enhancing group. Combining these observations, a semi-synthetic design of triterpene compound 54 (FIG. 9) which was 500-fold more active than any other known triterpene compound in inhibiting the inflammatory enzymes iNOS (nitric oxide synthase) and COX-2 (cyclooxygenase-2) (FIG. 9). Synthesis of Compounds of formulae (II), (IIA) and (IIB)
The 13-step synthesis of formula (II), (IIA) and (IIB) (as shown in figures 4 and 8, synthesis schemes 1 and 4, respectively) is efficient and enables the preparation of various analogs for SAR studies. The biological importance of the unique tricyclic skeletons of the compounds of formula (II), (IIA) and (IIB) (C8 epimer was constructed with a suitable Diels-Alder catalyst). The sites susceptible to alteration by the synthetic methods of the invention or by standard modifications of our synthetic intermediates are shown in FIG. 10, and representative examples of compounds of formula (II) are shown in FIG. 11.
The desired chemical backbone of the compounds of formula (II), (IIA) and (IIB) can also be incorporated onto a solid support such as Wang resin. This allows the easy construction of combinatorial libraries of compounds of formula (II), (IIA) and (IIB). In addition, preferred TNF-alpha and IL-1 modulators currently available can be identified and screened more rapidly in accordance with the present invention.
Plain and labeling experiments. It is also preferred that the backbone of the compounds of formula (II), (IIA) and (IIB) is labelled with a reactive cross-linker useful in photoaffinity labelling assays. These experiments help to determine the in vivo targets of the compounds of formulae (II), (IIA) and (IIB) and provide a basic understanding of the mode of action of the aconthoic acid and the activation of TNF- α. C19 carboxylic acid or C15 aldehyde (precursor of 1) can be used to perform cross-linking experiments with appropriate photosensitizing agents (see 60 and 61, FIG. 12). Synthesis of dimers and conjugates of Compounds of formulae (II), (IIA) and (IIB)
Dimeric forms of the compounds of formula (II), (IIA) and (IIB) have been isolated from natural sources such as 62(n ═ 1), and in addition, dexamethasone-antanthoic acid conjugates 63 provide interesting biological consequences, i.e. the goal is to obtain drugs that target steroid receptors, which are of potential value in cancer research. See chammy, m.c.; piowano, m.; garbarino, j.a.; miranda, c.; vicente, g. phytochemistry 1990, 9, 2943-2946. Although no biological studies have been conducted on this class of compounds, dimeric analogues of formula (II), (IIA) and (IIB) were evaluated according to the invention. The biologically active analogs of synthetic acoNToic acid or 1 are used as monomeric partners and their coupling is carried out using standard techniques, including those described herein. Experimental techniques
All reactions were carried out under argon atmosphere in dry freshly distilled solvent under anhydrous conditions unless otherwise indicated. Tetrahydrofuran (THF) and diethyl ether (Et)2O) is distilled from sodium/benzophenone; dichloromethane (CH)2Cl2) Hexamethylphosphoramide (HMPA), and toluene were distilled from calcium hydride; dimethylformamide (DMF) was distilled from calcium chloride. Unless otherwise indicated, product refers to chromatography and spectroscopy: (1H NMR) homogeneous material. Unless otherwise indicated, reagents were purchased in top grade commercial quality and used without further purification. The reaction was monitored by thin layer chromatography on a 0.25mm e.merck silica gel plate (60F-254) using UV light as developing agent, 7% ethanolic phosphomolybdic acid or p-anisaldehyde solution and heating as developing agent. Flash chromatography purification was performed using E.Merck silica gel (60, particles 0.040-0.063 mm). Preparative thin layer chromatography was performed on 0.25 or 0.50mm E.Merck silica gel (60F-254). NMR spectra were recorded on a Varian 400 and/or 500Mhz apparatusRecorded and calibrated using residual non-deuterated solvent as an internal standard. The following abbreviations are used to explain the multiplicities: s is singlet; d is bimodal; t is a triplet; q is quartet, m is multiplet; b is broad peak. The IR spectra were recorded on a Nicolet Avatar 320 FT-IR spectrophotometer. Optical rotations were recorded on a Perkin Elmer 241 polarimeter. High Resolution Mass Spectra (HRMS) were recorded under Chemical Ionization (CI) conditions on a VG 7070 HS mass spectrometer or under fast atom bombardment ion (FAB) conditions on a VGZAB-ZSE mass spectrometer.Triketone 2. A solution of diketone 1(50g, 0.40mol) in ethyl acetate (500ml) was treated with triethylamine (72ml, 0.52mol) and methyl vinyl ketone (36ml, 0.44 mol). The reaction mixture was refluxed at 70 ℃ for 10 hours and then cooled to 25 ℃. The solvent was removed under reduced pressure and the crude product was directly chromatographed (10-40% diethyl ether in hexanes) to give triketone 2(61g, 0.31mol, 78%). 2: a colorless oil; rf0.25 (silicic acid, 50% diethyl ether in hexanes);
1HN MR(400MHz，CDCl3)δ2.75-2.59(m，4H)，2.34(t，2H，J＝7.2Hz)，2.10(s，3H)，2.07-2.05(m，3H)，1.98-1.94(m，1H)，1.24(s，3H).Wieland-Miescher Ketone (3) A solution of triketone 2(61g, 0.31mol) in dimethyl sulfoxide (400ml) was treated with finely ground D-proline (1.7g, 0.01 mol). The solution was stirred at 25 ℃ for 4 days and then at 40 ℃ for another 1 day. The resulting purple solution was cooled to 25 ℃, diluted with water (300ml) and brine (100ml) and poured into a separatory funnel. The mixture was extracted with diethyl ether (3X 800 ml). The organic layer was concentrated (not dried) and chromatographed (10-40% diethyl ether in hexanes) to give 59g of crude reddish-violet oil. The material was further chromatographed (10-40% diethyl ether in hexanes) and concentrated,57g of a yellow oil are obtained. The oil was dissolved in diethyl ether (400ml) and held at 4 ℃ for 30 minutes, then a layer of hexane (100ml) was added to the top of the diethyl ether. A small amount of crystals was added to the bilayer solution and placed in a refrigerator (-28 ℃ C.) and kept overnight. The resulting crystals were collected by filtration, washed with ice-cold hexane (2X 100ml), and dried under reduced pressure. The mother liquor was concentrated to give another crop of product, and the resulting crystals were combined to give Wieland-Miescher ketone (3) (43g, 0.24mol, 78%). 3: brown crystals; rf0.25 (silica gel, 50% diethyl ether in hexanes);
[α]25 D：-80.0(c＝1，C6H6)；1H NMR(400MHz，CDCl3)δ5.85(s，1H)，2.72-2.66(m，2H)，2.51-2.42(m，4H)，2.14-2.10(m，3H)，1.71-1.68(m，1H)，1.44(s，3H)：13C NMR(100MHz，CDCl3)δ210.7，198.0，165.6，125.7，50.6，37.7，33.7，31.8，29.7，23.4，23.0.acetal 4. A solution of ketone 3(43g, 0.24mol) in benzene (700ml) was treated with p-toluenesulfonic acid (4.6g, 0.024mol) and ethylene glycol (15ml, 0.27 mol). The reaction mixture was refluxed at 120 ℃ using a dean-stark trap and a condenser. Once no water was collected in the dean-stark trap, the reaction was complete (approximately 4 hours). The longer the reaction time, the darker the reaction mixture and the lower the overall yield. The reaction mixture was cooled to 25 ℃, quenched with triethylamine (5ml, 0.036mol) and poured into a separatory funnel containing water (300ml) and saturated sodium bicarbonate (200 ml). The resulting mixture was then extracted with diethyl ether (3X 800 ml). The combined organic layers were dried over magnesium sulfate, concentrated, and chromatographed (10-40% diethyl ether in hexanes) to give acetal 4(48g, 0.22mol, 90%). 4: a yellow oil; rf0.30 (silica gel, 50% diethyl ether in hexanes);
[α]25 D：-77(c＝1，C6H6) (ii) a IR (film coating) vmax 2943，2790，1667，1450，1325，1250；1H NMR(400MHz，CDCl3)d5.80(s，1H)，3.98-3.93(m，4H)，2.43-2.35(m，3H)， 2.34-2.20(m，3H)，1.94-1.82(m，1H)，1.78-1.60(m，3H)， 1.34(s，3H)；13C NMR(100MHz，CDCl3)δ198.9，167.5，125.5，112.2，65.4，65.1，45.1，34.0，31.5，30.1，26.9，21.8，20.6.Ketone ester 5. to a solution of lithium (0.72g, 0.10mol) in liquid ammonia (400ml) was added dropwise a solution of acetal 4(10g, 0.045mol) and tert-butanol (3.7ml, 0.045mol) in diethyl ether (40ml) at-78 ℃. The resulting blue mixture was warmed and stirred under reflux (-33 ℃) for 15 minutes, then cooled to-78 ℃. Sufficient isoprene (about 8ml) was added dropwise to eliminate the residual blue color of the reaction mixture. The reaction mixture was then warmed in a water bath (50 ℃) and the ammonia was evaporated rapidly under a stream of dry nitrogen. The remaining ether was removed under reduced pressure to give a white foam. After 5 minutes under high vacuum, the nitrogen atmosphere was returned, the lithium enol was suspended in anhydrous ether (150ml) and cooled to-78 ℃. Methyl cyanoformate (4.0ml, 0.050mol) was then added and the reaction stirred at-78 ℃ for 40 minutes. The reaction was warmed to 0 ℃ and stirred for an additional 1 hour. Water (300ml) and diethyl ether (200ml) were added, and the mixture was poured into a separatory funnel containing saturated sodium chloride (100 ml). After separating the organic layer, the aqueous layer was extracted with diethyl ether (2X 400 ml). The combined organic layers were dried over magnesium sulfate, concentrated, and purified by chromatography (10-40% diethyl ether in hexanes) to afford ketoester 5(7.0g, 0.025mol, 55%). 5: white powdery precipitate; rf0.40 (silica gel, 50% diethyl ether in hexanes);
[α]25 D：-2.9(c＝1，C6H6) (ii) a IR (film coating) vmax 2943，1746，1700；1HNMR(400MHz，CDCl3)δ4.00-3.96(m，2H)，3.95-3.86(m，2H)，3.74(s，3H)，3.23(d，1H，J＝13.2Hz)，2.50-2.42(m，3H)，2.05-1.92(m，1H)，1.79-1.50(m，5H)，1.32-1.28(m，2H)，1.21(s，3H)；13CNMR(100MHz，CDCl3)δ205.4，170.0，111.9，65.2，65.1，59.9，52.0，43.7，41.6，37.5，30.3，29.8，26.2，22.5，14.0；HRMS，C15H22O5(M+Na+) Calculated value 305.1359 of (g), found value 305.1354.Ester 6. A solution of ketoester 5(7.0g, 0.025mol) in HMPA (50ml) was treated with sodium hydride (0.71g, 0.030 mol). After stirring for 3 hours at 25 ℃ the resulting yellow-brown reaction mixture was treated with chloromethyl methyl ether (2.3ml, 0.030mol) and the reaction was stirred for a further 2 hours at 25 ℃. The resulting white-yellow mixture was then poured into a separatory funnel containing ice water (100ml), saturated sodium bicarbonate (50ml), and diethyl ether (200 ml). After separation of the layers, the aqueous layer was extracted with ether (3X 200 ml). The combined ether extracts were dried over magnesium sulfate, concentrated, and purified by chromatography (silica gel, 10-40% diethyl ether in hexanes) to give ester 6(7.7g, 0.024mol, 95%). 6: a yellow oil; rf0.45 (silica gel, 50% diethyl ether in hexanes);
[α]25 D：+26.3(c＝1，C6H6) (ii) a IR (film coating) vmax 2951，1728，1690，1430，1170；1HNMR(400MHz，CDCl3)δ4.89(dd，2H，J＝22.8，6.4Hz)，3.93-3.91(m，2H)，3.90-3.84(m，2H)，3.69(s，3H)，3.40(s，3H)，2.72-2.68(m，1H)，2.24(bs，2H)，1.80-1.42(m，4H)，1.37-1.15(m，2H)，0.960(s，3H)，0.95-0.80(m，2H)；13C NMR(100MHz，CDCl3)δ167.8，150.5，115.8，112.1，93.0，65.2，65.1，56.3，51.3，40.7，40.3，30.3，26.4，23.6，22.9，22.3，13.9；HRMS，C17H26O6(M+Na+) Calculated value 349.1622 of (g), found value 349.1621.Aldol 7. addition of lithium (1.1g, 0.10mol) to liquid ammonia (1.1g, 0.10mol) at-78 ℃400ml) of the mixture was added dropwise a solution of the ester 6(7.7g, 0.024mol) in 1, 2-DME (30 ml). The blue reaction mixture was warmed and stirred under reflux (-33 ℃) for 20 minutes. The reaction mixture was then cooled to-78 ℃ and rapidly treated with excess methyl iodide (15ml, 0.24 mol). The resulting white slurry was stirred under reflux (-33 ℃) for 1 hour, then the reaction was warmed in a water bath (50 ℃) to allow the ammonia to evaporate. The reaction mixture was treated with water (100ml), sodium bicarbonate (100ml), and diethyl ether (200ml), and poured into a separatory funnel. After separation of the layers, the aqueous layer was extracted with ether (3X 200 ml). The combined ether extracts were dried over magnesium sulfate, concentrated, and chromatographed (silica gel, 10-30% diethyl ether in hexanes) to give acetal 7(4.1g, 0.014mol, 61%). 7: a semi-crystalline yellow oil; rf0.80 (silica gel, 50% diethyl ether in hexanes);
[α]25 D：+16.9(c＝10，C6H6) (ii) a IR (film coating) vmax 2934，1728，1466，1379，1283，1125，942；1H NMR(400MHz，CDCl3)δ3.95-3.80(m，4H)，3.64(s，3H)， 2.17-2.15(m，1H)，1.84-1.37(m，11H)，1.16(s，3H)，1.05-1.00(m，1H)，0.87(s，3H)；13C NMR(100MHz，CDCl3)δ177.7，112.9，65.2，64.9，51.2，44.0，43.7，38.1，30.7，30.3，28.8，23.4，19.1，14.7；HRMS，C16H26O4(M+H+) Calculated value 283.1904 of (g), found value 283.1904.Ketone 8 to a solution of acetal 7(4.1g, 0.014mol) in THF (50ml) was added dropwise 1M HCl (ca. 15ml) with stirring at 25 ℃. The reaction was monitored by thin layer chromatography and once the starting material disappeared, it was neutralized with sodium bicarbonate (30 ml). The resulting mixture was poured into a separatory funnel containing water (100ml) and diethyl ether (100 ml). After separation of the layers, the aqueous layer was extracted with ether (3X 100 ml). The combined ether extracts were dried over magnesium sulfate and concentratedCondensation and chromatographic purification (silica gel, 10-20% diethyl ether in hexane) gave ketone 8(3.3g, 0.014mol, 95%). 8: white crystals; rf0.70 (silica gel, 50% diethyl ether in hexanes);
[α]25 D：+3.5(c＝1.0，C6H6) (ii) a IR (film coating) vmax 2943，1728，1449，1239，1143，1095，985；1H NMR(400MHz，CDCl3)δ3.62(s，3H)，2.55-2.45(m，1H)，2.92-1.95(m，5H)，1.8-1.6(m，2H)，1.50-1.30(m，4H)，1.14(s，3H)，0.98-0.96(m，1H)，0.90(s，3H)，13C NMR(100MHz，CDCl3)δ214.8，177.0，54.4，51.3，49.3，44.2，37.9，37.7，33.1，28.6，26.4，22.8，18.8，17.0；HRMS，C14H22O3(M+Na+) Calculated value 261.1461 of (g), found value 261.1482.Alkyne 9. A solution of ketone 8(2.0g, 8.3mmol) in diethyl ether (50ml) was treated with lithium acetylide (0.40g, 13 mmol). The reaction was stirred at 25 ℃ for 1 hour, then quenched with sodium bicarbonate (20ml) and water (30 ml). The mixture was poured into a separatory funnel and the layers were separated. The aqueous layer was extracted with diethyl ether (3X 50 ml). The combined organic layers were dried over magnesium sulfate, concentrated, and chromatographed (silica gel, 10-30% diethyl ether in hexanes) to give alkyne 9(2.0g, 7.6mmol, 90%). 9: a white solid; rf0.65 (silica gel, 50% diethyl ether in hexanes);
1HNMR(400MHz，CDCl3)δ3.64(s，3H)，2.56(s，1H)，2.18-2.10(m，1H)，1.92-1.40(m，12H)，1.18(s，3H)，1.17-1.01(m，1H)，0.81(s，3H)；13C NMR(100MHz，CDCl3)177.6，86.8，76.5，75.0，51.2，50.5，43.9，52.5，37.9，35.3，33.4，28.8，23.5，22.5，19.1，11.5；HRMS，C16H24O3(M+H+-H2o) calculated 247.1693, found 247.1697.Alkene 10. A solution of alkyne 9(0.50g, 1.9mmol) in 1, 4-dioxane (20ml) and pyridine (2ml) was treated with Lindele catalyst (100 mg). The mixture was placed under pressure (30 lbs/in)2) Hydrogenation was carried out for 7 minutes. The reaction mixture was then diluted with ether (10ml), filtered through a pad of celite and washed with ether (2 × 50 ml). The solvent was evaporated under reduced pressure to give olefin 10(0.48g, 1.8mmol, 95%). 10: a colorless oil;
1H NMR(400MHz，CDCl3)δ6.58(dd，1H)，5.39(d，1H)，5.14(d，1H)，3.64(s，3H)，2.20-2.11(m，2H)，1.93-1.65(m，4H)，1.61(s，2H)，1.52-1.25(m，4H)，1.19(s，3H)，1.17-0.90(m.2H)，0.89(s，3H).diene 11. A solution of olefin 10(0.48g, 1.8mmol) in benzene (80ml) and THF (20ml) was treated with boron trifluoride etherate (1ml, 7.9mmol) and the reaction mixture was refluxed at 100 ℃ for 5 hours. After cooling, the reaction was treated with 1N NaOH (1ml, 26mmol) and the mixture was poured into a separatory funnel containing water (100ml) and diethyl ether (100 ml). After separation of the layers, the aqueous layer was extracted with ether (3 × 100ml), the organic layers were combined, dried over magnesium sulfate, concentrated, and chromatographed (silica gel, 5% ether in hexane) to give diene 11(0.42g, 1.7mmol, 95%), 11: a colorless oil; rf0.95 (silica gel, 50% diethyl ether in hexanes);
1H NMR(400MHz，CDCl3)δ6.26-6.23(dd，1H)，5.70(s，1H)，5.253(d，1H，J＝19.2Hz)，4.91(d，1H，J＝12.8Hz)，3.64(s，3H)，2.22-2.12(m，2H)，2.10-1.94(m，2H)1.92-1.67(m，3H)，1.60-1.44(m，3H)，1.378(d，1H，J＝13.6)，1.21(s，1H)，1.19-1.00(m，2H)，0.86(s，3H)；13C NMR(100MHz，CDCl3)δ177.7，146.7，136.1，121.9，113.3，53.0，51.2，43.9，38.0，37.9，37.4，28.5，27.8，20.5，19.5，18.3.aldehyde 12A solution of methacrolein (0.5ml, 5.2mmol) and diene 11(0.1g, 0.40mmol) was stirred neat at 25 ℃ for 8 h. Excess methacrolein was then removed under reduced pressure. The crude product was chromatographed (silica gel, 10-20% diethyl ether in hexanes) to afford aldehydes 12 and 12*(0.13g, 0.40mmol, 100%) as a mixture of diastereomers at a ratio of 3: 1 to 4: 1 at C13. 12 and 12*: a colorless oil; rf0.55 (silica gel, 25% diethyl ether in hexanes);
12: IR (film coating) vmax 3441，2936，1726，1451，1233，1152：1H NMR(400MHz，CDCl3)δ9.70(s，1H)，5.58(m，1H)，3.62(s，3H)，2.38-2.25(m，1H)，2.21-2.18(m，1H)，2.17-1.98(m，4H)，1.96-1.62(m，6H)，1.61-1.58(m，1H)，1.57-1.43(m，2H)，1.40-1.23(m，1H)，1.17(s，3H)，1.04(s，3H)，0.92(s，3H)；13C(100MHz，CDCl3)δ207.6，177.7，148.3，188.6，51.3，47.8，47.0，44.2，41.2，39.3，38.8，38.1，29.5，28.4，22.9，22.5，21.8，20.6，20.5，19.7；12*：[α]25 D：+36.8(c＝0.7，C6H6) (ii) a IR (film coating) vmax 3441，2936，1726，1451，1233，1152；1H NMR(400MHz，CDCl3)δ9.64(s，1H)，5.42(m，1H).3.66(s，3H)，2.29-2.10(m，4H)，2.09-1.84(m，4H)，1.81-1.77(m，2H)，1.75-1.63(m，2H)，1.62-1.58(m，2H)，1.57-1.45(m，1H)，1.43(s，1H)，1.13(s，3H)，1.03(s，3H)，0.87(s，3H)；13C NMR(100 MHz，CDCl3)δ207.3，177.5，147.4，114.6，55.8，51.3，47.3，44.5，40.7，40.4，38.4，37.5，3 1.5，28.6，25.0，24.2，21.9，19.9，19.6，18.7.
The preferred method for purifying the diastereomeric aldehyde is reduction with sodium borohydride in MeOH and separation of the alcohol. The main compound (diastereomer from this end up) may then be oxidized by treatment with Dess-Martin periodinane to give the desired aldehyde 12.Olefin 13(TTL3) a solution of (methyl) -triphenyl-phosphonium bromide (357mg, 1.0mmol) in THF (40ml) was treated with a 1M solution of NaHMDS in THF (0.86ml, 0.86 mmol). The resulting yellow mixture was stirred at 25 ℃ for 30 minutes. A solution of aldehyde 12(91mg, 0.29mmol) in THF (10ml) was then added to the reaction via a cannula. The reaction mixture was stirred at 25 ℃ for 8 hours, and then quenched with sodium bicarbonate (30ml) and water (20 ml). The mixture was poured into a separatory funnel containing diethyl ether (50 ml). After separation of the layers, the aqueous layer was extracted with ether (3X 50 ml). The combined organic layers were dried over magnesium sulfate, concentrated, and purified by chromatography (silica gel, 10% diethyl ether in hexanes) to give alkene 13(84mg, 0.28mmol, 97%). 13: a colorless oil; rf0.75 (silica gel, 25% diethyl ether in hexanes);
13：1HNMR(400MHz，CDCl3)δ5.96(dd，1H，J＝16.8，11.6Hz)，5.50(m，1H)，4.98(m，2H)，3.62(s，3H)，2.20-2.11(m，1H)，2.10-1.91(m，4H)，1.90-1.70(m，4H)，1.69-1.51(m，3H)，1.50-1.38(m，3H)，1.36-1.24(m，1H)，1.17(s，3H)，1.04(s，3H)，0.90(s，3H)；13C NMR(100MHz，CDCl3)δ177.9，149.1，143.8，117.9，111.7，51.2，47.7，44.4，41.4，41.2，38.9，38.3，37.7，34.8，30.4，28.4，24.8，23.1，22.3，22.2，20.6，19.8.acid 14(TTL1) a solution of alkene 13(84mg, 0.28mmol) in dimethyl sulfoxide (20ml) was treated with LiBr (121mg, 1.4 mmol). The reaction mixture was refluxed at 180 ℃ for 2 days. After cooling, the reaction was diluted with water (30ml) and extracted with diethyl ether (3X 50 ml). The combined organic layers were dried over magnesium sulfate, concentrated, and purified by chromatography (silica gel, 30% diethyl ether in hexanes) to afford carboxylic acid 14(TTL1) (78mg, 0.26 mmol). 14: a white solid; rf0.30 (silica gel, 30% diethyl ether in hexanes);
1H NMR(400MHz，CDCl3)δ5.96(dd，1H，J＝14.4，9.6Hz)，5.52(m，1H)，4.98-4.95(m，2H)，2.20-1.72(m，10H)，1.64-1.58(m，3H)，1.57-1.37(m，4H)，1.22(s，3H)，1.04(s，3H)，0.99(s，3H)；13C NMR(100MHz，CDCl3) Delta 182.9, 149.3, 143.9, 118.1, 111.9, 47.5, 44.2, 41.3, 41.2, 38.9, 38.0, 37.6.34.8, 28.4, 24.7, 23.0, 22.4, 21.9, 20.3, 19.53P＝14CH2.
Triphenylphosphine (0.16g, 0.61mmol) was added to a 15ml reaction flask and dried under vacuum at 25 ℃ overnight. To the flask was added 2ml THF (dried and degassed under vacuum), then 1ml THF dissolved14CH3I (50mCi, 53mCi/mmol, 0.9mmol) and the mixture was stirred under argon for 24 h. Potassium hexamethyldisilylamide (2.5ml, 1.25mmol, 0.5M in toluene) was then added and the reddish-yellow mixture was stirred at 25 ℃ for 3 hours. By Ph3P＝14CH2Carrying out the wittig reaction
The mixture was cooled at-78 ℃ and treated with aldehyde 12(63mg, 0.2mmol) in dry THF (1.5 ml). The mixture was slowly warmed to 25 ℃, stirred for 8 hours, and quenched with sodium bicarbonate (10ml) and water (10 ml). The mixture was extracted with diethyl ether (3X 50ml), the organic layers combined and washed with sulphurMagnesium was dried, concentrated and chromatographed on silica gel (silica gel, 10% diethyl ether in hexanes) to give the olefin 13.Alcohol 15. A solution of alkyne 9(1.10g, 4.2mmol), thiophenol (1.37g, 12.4mmol) and 2, 2' -azobisisobutyronitrile (AIBN, 34.5mg, 0.21mmol) in xylene (25ml) was stirred at 110 deg.C (under argon) for 18 h. The reaction mixture was cooled to 25 ℃ and quenched with saturated aqueous sodium bicarbonate (50 ml). The organic layers were extracted with ether (3X 50ml), combined and dried (MgSO)4) Concentrated and the residue chromatographed (silica gel, 2-5% diethyl ether in hexanes) to give alcohol 15(1.35g, 3.6mmol, 85.7%); 15: a colorless liquid; rf0.51 (silica gel, 5% diethyl ether in hexanes);
[α]25 D: +24.20(c ═ 1.0, benzene); IR (film coating) vmax 2946.8，1724.5，1472.6，1438.4，1153.5，740.0，690.9；1H NMR(500MHz，CDCl3)δ7.20-7.60(m，5H)，5.23(d，1H，J＝10.5Hz)，5.12(d，1H，J＝10.0Hz)，3.62(s，3H)，2.08-2.24(m，2H)，1.16-1.92(m，9H)，1.09(s，3H)，0.86-1.02(m，2H)，0.68(s，3H)；13C NMR(100MHz，CDCl3)δ177.8，151.7，133.9，133.7，128.8，127.9，118.2，54.9，53.5，51.1，44.3，40.4，38.1，37.3，28.7，27.7，25.5，23.5，19.5，18.5.Diene 16. to a solution of alcohol 15(1.10g, 2.94mmol) in hexamethylphosphoramide (HMPA, 10ml) was added phosphorus oxychloride (0.50g, 3.3mmol) dropwise and the mixture was stirred at 25 ℃ until clear. Pyridine (0.26mol, 3.23mmol) was then added and the mixture was stirred at 150 deg.C (under argon) for 18 hours. The reaction mixture was cooled to 25 ℃ and quenched with saturated aqueous sodium bicarbonate (50 ml). The organic layers were extracted with ether (3X 60ml), combined and dried (MgSO)4) Concentrated and the residue chromatographed (silica gel, 2-5% diethyl ether in hexane) to yieldDiene 16(0.85g, 2.38mmol, 81%); 16: a colorless liquid; rf0.60 (silica gel, 5% diethyl ether in hexanes);
[α]25 D: -17.30(c ═ 1.08, benzene); IR (film coating) vmax2957.0，1726.6，1581.6，1478.3，1439.0，1234.7，1190.8，1094.8，1024.4，739.1；1H NMR(500MHz，CDCl3)δ7.20-7.60(m，5H)，6.43(d，1H，J＝15.0Hz)，6.36(d，1H，J＝14.5Hz)，5.72(m，1H)，3.64(s，3H)，1.48-2.32(m，10H)，1.43(s，3H)，1.21(s，3H)，1.05(m，1H)，0.88(s，3H)；13C NMR(125MHz，CDCl3)δ177.9，133.7，129.1，128.9，128.6，127.5，126.2，123.4，120.9，52.8，51.1，43.7，37.7，37.3，30.2，28.3，27.7，20.1，19.3，18.3.Aldehyde 17 to a solution of diene 16(0.51g, 1.43mmol) and methacrolein (0.30g, 4.30mmol) in dichloromethane (5ml) was added dropwise tin (IV) chloride (0.29ml of a 1M solution in dichloromethane, 0.29mmol) at-20 ℃ under argon. The resulting mixture was warmed to 0 ℃ over 1 hour and stirred at 0 ℃ for 18 hours. The reaction was quenched with saturated aqueous sodium bicarbonate (15ml) and the organic layer was extracted with diethyl ether (3X 20 ml). The combined organic layers were dried (MgSO)4) Concentrated and the residue chromatographed (silica gel, 10-15% diethyl ether in hexanes) to give aldehyde 17(0.51g, 1.19mmol, 83.7%); 4: a colorless liquid; rf0.48 (silica gel, 10% diethyl ether in hexanes);
[α]25 D: +30.0(c ═ 1.13, benzene); IR (film coating) vmax 2930.8，2871.4，1724.9，1458.4，1226.4，1149.8；1H NMR(500MHz，CDCl3)δ9.51(s，1H)，7.20-7.60(m，5H)，5.57(m，1H)，3.65(s，3H)，1.20-2.32(m，15H)，1.17(s.3H)，1.05(s，3H)，0.91(s.3H)：13C NMR(125MHz，CDCl3)δ203.6，177.9，153.7，133.6，133.5，128.9，127.8，117.1，51.3，49.1，47.7，44.2，41.6，38.7，38.1，31.2，28.3，27.8，26.9，21.7，20.2，19.3，18.6.Alcohol 18 to a solution of aldehyde 17(0.50g, 1.17mmol) in anhydrous ethanol (5ml) was added sodium borohydride (50mg, 1.32mmol) portionwise and the mixture was stirred for 30 min. Saturated aqueous sodium bicarbonate (10ml) was then added and the mixture was extracted with diethyl ether (3X 20 ml). The combined organic layers were dried (MgSO)4) And concentrated. The residue was dissolved in tetrahydrofuran (5ml) and treated with Raney nickel at 65 ℃ for 10 minutes under an argon atmosphere. The reaction mixture was filtered and the filtrate was dried (MgSO4) And concentrated, and the residue chromatographed (silica gel, 2-5% diethyl ether in hexanes) to give alcohol 18 as the major compound (0.21g, 0.65mmol, 56.1% overall yield). Note: the total yield of the two reactions is 91%); 18: a colorless liquid; rf0.39 (silica gel, 30% diethyl ether in hexanes);
[α]25 D：-6.70(c＝1.0，
benzene); IR (film coating) vmax 3436.8，2929.0，2872.2，1728.1，1433.9，1260.6，1029.7，801.6；1H NMR(500MHz，CDCl3)δ5.37(m，1H)，3.62(s，3H)，2.28(bs，1H)，2.06-2.20(m，2H)，1.20-2.00(m，12H)，1.16(s，3H)，0.99(m，1H)，0.86(s，3H)，0.84(s，3H)；13C NMR(125MHz，CDCl3)δ178.2，150.4，16.4，73.6，51.2，47.9，44.2，41.9，38.8，38.2，34.3，33.9，28.3，28.2，27.8，22.1，20.3，18.9.Olefin 19 to a solution of alcohol 18(20.0mg.0.062mmol) in dichloromethane (2ml) Dess-Martin periodinane (35mg, 0.08mmol) was added in portions and the mixture was stirred at 25 ℃ for 30 minutes. The reaction was terminated with saturated aqueous sodium bicarbonate (5ml)The reaction was stopped and extracted with diethyl ether (3X 10 ml). The combined organic phases were dried (MgSO)4) And concentrated. The residue was redissolved in tetrahydrofuran (0.5ml) and added to a yellow suspension of (methyl) triphenylphosphonium bromide (60mg, 0.17mmol) and sodium bis (trimethylsilyl) amide (0.14ml of a 1.0M solution in THF) in THF (1.5ml) under an argon atmosphere. After stirring at 25 ℃ for 18 h, the mixture was diluted with saturated aqueous sodium bicarbonate (5ml) and extracted with diethyl ether (3X 10 ml). The combined organic layers were dried (MgSO)4) Concentration and chromatography of the residue (silica gel, 2-5% diethyl ether in hexane) gave olefin 19(16.8mg, 0.05mmol, 86% overall yield of the two steps); 19: a colorless liquid; rf0.74 (silica gel, 5% diethyl ether in hexanes);
[α]25 D：-14.40
(c ═ 0.50, benzene); IR (film coating) vmax 2929.5，2873.4，1726.8，1637.7，1460.7，1376.8，1225.1，1150.4，997.8，908.7；1H NMR(500MHz，CDCl3)δ5.82(dd，1H)，5.39(m，1H)，4.85-4.94(dd，2H)，3.64(s，3H)，2.30(bs，1H)，2.14(m，1H)，2.02(m，1H)，1.80-1.98(m，2H)，1.68-1.80(m，2H)，1.20-1.68(m，7H)，1.18(s，3H)，0.96-1.08(m，2H)，0.95(s，3H)，0.88(s，3H)；13C NMR(125MHz，CDCl3)δ178.3，150.4，125.6，116.6，109.2，51.2，47.9，44.3，41.9，41.8，38.3，38.2，37.4，34.8，30.2，29.6，28.6，28.4，27.8，22.1，20.4，19.0.To a solution of alkene 19(16.8mg, 0.05mmol) in N, N-dimethylformamide (2ml) was added lithium bromide (5.0mg, 0.06mmol) and the mixture was refluxed at 190 ℃ for 1 hour. The reaction mixture was then cooled to 25 ℃, diluted with water (5ml) and extracted with ethyl acetate (3 × 10 ml). The organic layers were combined and dried (Mg)SO4) Concentrated and the residue chromatographed (silica gel, 15-20% diethyl ether in hexanes) to give formula (I) (14.9mg, 0.05mmol, 92.6%); the compound of formula (I) is a colorless liquid; rf0.20 (silica gel, 30% diethyl ether in hexanes);
[α]25 D：
-6.0(c ═ 0.33, benzene); IR (film coating) vmax 3080.6，2928.9，2857.6，1693.6，1638.2，1464.7，1413.8，1376.4，1263.1，1179.3，1095.9，1027.5，999.2，909.2，801.7；1H NMR(500MHz，CDCl3)δ5.82(dd，，1H)，5.40(m，1H)，4.85-4.95(dd，2H)，2.30(bs，1H)，2.16(m，1H)，2.02(m，1H)，1.80-1.98(m，2H)，1.70-1.84(m，2H)，1.10-1.70(m，7H)，1.24(s，3H)，1.00-1.10(m，2H)，0.99(s，3H)，0.95(s，3H)；13CNMR(125MHz，CDCl3) Delta 150.3, 149.9, 116.7, 109.2, 47.9, 41.8, 41.7, 38.3, 38.2, 37.4, 34.8, 31.8, 28.6, 28.5, 27.7, 22.6, 22.4, 22.1, 20.3, 18.9 using the process of the invention
The in vitro and in vivo methods described hereinabove as part of the present invention also establish the selectivity of TNF-alpha or IL-1 modulators. It will be appreciated that the compounds may modulate a wide variety of biological processes, or may be selective. The specificity of candidate modulators can be determined using cell plates based on the invention. Selectivity is evident in, for example, the field of chemotherapy, where selectivity for compounds that are toxic to cancer cells, but not to non-cancer cells, is clearly desirable. Selective modulators are preferred because they have fewer side effects in a clinical setting. The selectivity of a candidate modulator can be determined in vitro by testing its toxicity and effect on a variety of cell lines exhibiting a variety of cellular pathways and sensitivities. The data obtained from these in vitro toxicity assays can be extended to animal models, including accepted animal model assays and human clinical assays, to determine the toxicity, efficacy, and selectivity of candidate modulators.
The invention also includes compositions made by the process of the invention, pharmaceutical compositions prepared for storage and administration comprising a pharmaceutically acceptable carrier comprising a pharmaceutically effective amount of the product disclosed above in a pharmaceutically acceptable carrier or diluent. Pharmaceutically acceptable carriers or diluents for use in therapy are well known in the Pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing co. (a.r. gennaro edge.1985). Preservatives, stabilizers, dyes and flavoring agents may be included in the pharmaceutical compositions. For example, sodium benzoate, ascorbic acid and parabens may be added as preservatives. In addition, antioxidants and suspending agents may be used.
These TNF- α or IL-1 modulator compositions may be formulated and used as tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions, suspensions for administration by injection; patches for transdermal administration, subcutaneous drug depot devices, and the like. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for formulation as solutions or suspensions in liquids prior to injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine hydrochloride and the like. In addition, the injectable pharmaceutical composition may contain small amounts of non-toxic excipients, such as wetting agents, pH buffering agents and the like, if desired. If desired, absorption-promoting agents (e.g., liposomes) can be used.
The pharmaceutically effective amount of the TNF- α or IL-1 modulator composition required as a dosing dose depends on the route of administration, the type of animal being treated, and the physical condition of the particular animal under consideration. The dosage can be adjusted to achieve the desired effect, but the dosage will depend on body weight, diet, concurrent medication, and other factors known to those skilled in the medical arts.
In practicing the methods of the invention, the products or compositions can be used alone or in combination with each other, or in combination with other therapeutic or diagnostic agents. These products can be used in vivo, generally in mammals, preferably in humans, or in vitro. When used in vivo, the products or compositions of the present invention may be administered to an animal in different dosage forms by different routes, including parenteral, intravenous, subcutaneous, intramuscular, colonic, rectal, vaginal, nasal, or intraperitoneal routes of administration. Such methods can also be used to test the activity of compounds in vivo.
It will be apparent to those skilled in the art that the dosage and particular mode of administration for in vivo administration will vary with the age, body weight, type of mammal being treated, the particular compound employed, and the particular application for which it is employed. Determination of an effective dosage level, i.e., the level of dosage required to achieve a desired result, can be accomplished by one skilled in the art using routine pharmacological methods. In general, for humans, the clinical use of the product is to start with a lower dosage level and then gradually increase the dosage level until the desired effect is achieved. Alternatively, acceptable in vitro assays can be used to determine suitable dosages and routes of administration for the compositions determined by the methods of the invention using established pharmacological methods.
In non-human animal studies, possible products are administered starting from higher dose levels and then gradually decreasing the dose until the desired effect is no longer achieved or the adverse side effects disappear. The dosage of the product of the invention may vary within wide limits, depending on the desired effect and the therapeutic indication. The dosage may generally be from about 10 micrograms/kg to 100mg/kg body weight, preferably from about 100 micrograms/kg to 10mg/kg body weight. Alternatively, the dose can be estimated and calculated from the surface area of the patient according to methods known to those skilled in the art. Administration is preferably carried out orally once daily or twice daily.
The appropriate formulation, route of administration and dosage may be selected by the particular practitioner according to the patient's condition. See, for example, Fingl et al, The Pharmacological Basis of Therapeutics, 1975. It should be noted that clinicians know how and when to discontinue, interrupt, or adjust dosing due to toxicity or organ dysfunction. Conversely, if the clinical response is inadequate (excluding toxicity), the clinician should also know how to adjust the treatment to higher levels. In the treatment of diseases, the dosage administered may vary with the severity of the condition being treated and the route of administration. The severity of the condition can be assessed, for example, in part, by standard diagnostic evaluation methods. In addition, the dosage and perhaps the frequency of administration will also vary with the age, weight, and response of the particular patient. Protocols comparable to those described above can be used in veterinary medicine.
Such therapeutic agents may be formulated and administered systemically or topically, depending on the particular condition being treated. Various techniques for formulation and administration can be found in Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing co., Easton, PA (1990). Suitable routes of administration may include oral, rectal, transdermal, vaginal, transmucosal, or intestinal administration; parenteral administration, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections.
For injection, the active agents of the invention may be formulated in aqueous solutions, preferably in physiologically acceptable buffers such as Hanks 'solution, ringer's solution, or physiological saline buffer. For such transmucosal administration, penetrants which promote a permeation barrier are used in the formulation. Such penetrants are generally known in the art. It is also within the scope of the present invention to formulate the compounds disclosed herein into dosage forms suitable for systemic administration using a pharmaceutically acceptable carrier. By proper selection of the carrier and appropriate manufacturing procedures, the compositions of the present invention, particularly those formulated as solutions, can be administered parenterally, for example by intravenous injection. The compounds of the present invention can be readily formulated into dosage forms suitable for oral administration using pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries (suspensions), suspensions and the like, for oral ingestion by a patient to be treated.
Active agents for intracellular administration can be administered using techniques well known to those skilled in the art. For example, such active agents may be encapsulated within liposomes and then administered as described above. All molecules present in the aqueous solution are incorporated into the aqueous interior when the liposomes are formed. The liposome contents are protected from the external microenvironment and, because the liposomes fuse with the cell membrane, they are efficiently delivered into the cell cytoplasm. In addition, small organic molecules can be administered directly intracellularly due to their hydrophobicity.
Pharmaceutical compositions suitable for use in accordance with the teachings herein include compositions which contain an effective amount of a TNF- α or IL-1 modulator for the purpose of TNF- α or IL-1 modulation. Determination of an effective amount is within the ability of those skilled in the art, especially in light of the detailed disclosure provided herein. In addition to the active ingredient, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations for pharmaceutical use. Formulations formulated for oral administration may be in the form of tablets, dragees, capsules or solutions. The pharmaceutical compositions of the present invention may be prepared in a manner that is itself known, for example, by means of conventional mixing, dissolving, granulating, dragee-making, levitating, emulsifying, encapsulating, or lyophilizing processes.
Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or other organic oils such as soybean oil, grape fruit oil, or almond oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol or dextran. The suspending agents may also optionally contain suitable stabilizers or agents that increase the solubility of the compounds to enable the preparation of highly concentrated solutions.
Pharmaceutical preparations for oral use can be prepared by: the active compound is mixed with a solid excipient, the resulting mixture is optionally milled, if necessary with suitable excipients, and the mixture of granules is then processed to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate may be added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions, lacquer solutions, and suitable organic solvents or solvent mixtures, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, can be used. Dyes or pigments may be added to the tablets or dragee coatings to identify or indicate different combinations of active compound doses. Such formulations can be made by methods known in the art (see, e.g., U.S. patent 5,733,888 (injectable compositions); 5,726,181 (poorly water soluble compounds); 5,707,641 (therapeutically active proteins or peptides); 5,667,809 (lipophilic active agents); 5,576,012 (solubilizing polymeric agents); 5,707,615 (antiviral agents); 5,683,676 (granular drugs); 5,654,286 (topical formulations); 5,688,529 (oral suspension agents); 5,445,829 (long release formulations); 5,653,987 (liquid formulations); 5,641,515 (controlled release formulations); and 5,601,845 (spherical formulations)).
The efficacy and toxicity of the compounds of the invention can be evaluated using known methods. For example, the toxicological properties of a particular compound of the invention or a subset of compounds of the invention sharing some chemical moieties (subset) may be determined by determining its toxicity in vitro on a cell line, e.g., a mammalian, preferably a human cell line. The results of these studies often predict toxicity in animals, such as mammals, or more particularly in humans. Alternatively, toxicity of a particular compound of the invention in animal models, such as mice, rats, rabbits, or monkeys, can be determined using known methods. The potency of a particular compound of the invention can be determined using methods known in the art, for example, in vitro methods, animal models, or human clinical trials. There are in vitro models known in the art that can be used for almost a variety of conditions, including those that can be alleviated by the invention, including cancer, cardiovascular disease, and various immune dysfunctions. Similarly, acceptable animal models can be used to determine the efficacy of a compound to treat these conditions. When selecting a model to determine efficacy, one of skill in the art can select an appropriate model, dosage, and route of administration, as well as experimental protocols, by guidance of the state of the art. Of course, human clinical trials can also be used to determine the efficacy of the compounds of the invention in humans.
When used as anti-inflammatory agents, anti-cancer agents, tumor growth inhibiting compounds, or as a means of treating cardiovascular disease, the compounds of formula (II), (IIA), and preferably (IIB) may be administered by oral or non-oral routes. When administered orally, it may be administered orally in capsules, tablets, granules, sprays, syrups, or other oral forms. When administered non-orally, it may be administered as an aqueous suspension, or an oil preparation or the like, or as drops, suppositories, ointments, salves or the like. When administered via injection, it can be administered subcutaneously, intraperitoneally, intravenously, intramuscularly, intradermally, and the like by injection. Similarly, it may be administered topically, rectally, or vaginally, as deemed appropriate by one skilled in the art, to provide optimal contact of the compounds of the invention with the tumor and thereby inhibit tumor growth. It may also be administered locally at the site of a tumor or other condition, locally before or after tumor resection, or as part of the treatment of the disease as is known in the art. Similarly, controlled release formulations, depot formulations, and infusion pumps may be used for administration.
When used as an anti-tumor agent or to treat any other of the above-identified conditions, the compounds of formulae (II) and (IIA), and preferably (IIB), may be administered to a human patient orally or non-orally at the following doses: from about 0.0007 mg/day to about 7000 mg/day of active ingredient, more preferably from about 0.07 mg/day to about 70 mg/day of active ingredient, preferably 1 time per day, or less preferably more than 2 times per day to about 10 times per day. Alternatively and also preferably, the compounds of the invention are preferably administered continuously at the indicated doses, for example continuously by intravenous drip. Thus, for a patient weighing 70 kilograms, the preferred daily dosage of active anti-tumor component is from about 0.0007 mg/kg/day to about 35 mg/kg/day, more preferably from 0.007 mg/kg/day to about 0.035 mg/kg/day. However, as will be appreciated by those skilled in the art, in some instances it may be desirable to administer the anti-tumor compounds of the present invention in amounts that exceed, or even far exceed, the preferred dosage ranges described above in order to effectively and actively treat a particular severe or lethal tumor.
For the preparation of the compound of formula (II), the compound of formula (IIA), or the compound of formula (IIB) as a tumor growth inhibitory or antiviral compound, known surfactants, excipients, lubricants, suspending agents, and pharmaceutically acceptable film-forming substances and coating adjuvants and the like can be used. Preferably, alcohols, esters, sulfated aliphatic alcohols, and the like can be used as the surfactant; sucrose, glucose, lactose, starch, microcrystalline cellulose, mannitol, light anhydrous silicate, magnesium aluminate, magnesium silicate aluminate (magnesium silicate aluminate), synthetic aluminum silicate, calcium carbonate, acid sodium carbonate, calcium hydrogen phosphate, carboxymethylcellulose calcium, or the like may be used as the excipient; magnesium stearate, talc, hardened oil, and the like can be used as a lubricant; coconut oil, olive oil, sesame oil, peanut oil, soybean oil may be used as a suspending agent or lubricant; cellulose acetate phthalate as a carbohydrate such as a derivative of cellulose or sugar, or a methyl acetate-methacrylate copolymer as a polyvinyl derivative may be used as a suspending agent; plasticizers such as phthalates and the like may be used as suspending agents. In addition to the preferred ingredients described above, sweeteners, flavoring agents, coloring agents, preservatives and the like may be added to the formulations for administration of the compounds of this invention, particularly when the compounds are administered orally.
When compounds of formula (II), formula (IIA), and/or formula (IIB) are used as a means of treating redness of the skin, the compounds can be formulated with pharmaceutically acceptable carriers for topical administration as ointments or salves.
When compounds of formula (II), formula (IIA) and/or formula (IIB) are used as biochemical test reagents as described above, the compounds of the invention may be dissolved in an organic or aqueous organic solvent or applied directly to any of the various cultured cell systems. Organic solvents that can be used include, for example, methanol, methyl sulfoxide, and the like. The formulation may be, for example, a powder, granules or other solid inhibitor, or a liquid inhibitor prepared using an organic solvent or an aqueous organic solvent. Although the preferred concentration of the compounds of the present invention for use as cell cycle inhibitors is generally from about 1 to about 100. mu.g/ml, the most suitable amount varies depending on the type of cell system being cultured and the purpose of use, and is understood by those skilled in the art. It may also be desirable or preferred by those skilled in the art in some applications to use amounts outside of the above-described ranges.
The invention also includes the composition of the compounds of formula (II), formula (IIA) and/or formula (IIB) in a pharmaceutical composition comprising a pharmaceutically acceptable carrier. Such compositions may be prepared for storage and administration. Pharmaceutically acceptable carriers or diluents for use in therapy are well known in the pharmaceutical art and are described, for example, in Remington's pharmaceutical Sciences, Mack Publishing co. (a.r. gennaro edge.1985). For example, such compositions may be formulated and used as tablets, capsules, or solutions for oral administration; suppositories for rectal or vaginal administration; sterile solutions or suspensions for administration by injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for formulation as solutions or suspensions in liquids prior to injection, or as emulsions. Suitable excipients include, but are not limited to, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine hydrochloride, and the like. In addition, the injectable pharmaceutical composition may contain small amounts of non-toxic excipients, such as wetting agents, pH buffering agents and the like, if desired. If desired, absorption-promoting agents (e.g., liposomes) can be used.
The pharmaceutically effective amount of the composition required as a dose depends on the route of administration, the type of animal being treated, and the physical condition of the particular animal in question. The dosage can be adjusted to achieve the desired effect, but the dosage will depend on body weight, diet, concurrent medication, and other factors known to those skilled in the medical arts.
As noted above, the products or compositions of the present invention may be used alone or in combination with each other, or in combination with other therapeutic or diagnostic agents. These products can be used in vivo or in vitro. The dosage and most useful mode of administration suitable will vary with the age, weight, and animal being treated, the particular compound employed, and the particular application for which the compositions are employed. In the treatment of diseases, the dosage administered may vary with the severity of the condition being treated and the route of administration, and depending on the condition and its severity, the compositions of the invention may be formulated and administered systemically or topically. Various techniques for formulation and administration can be found in Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing co., Easton, PA (1990).
Various references, publications and patents are cited herein. To the extent allowed by law, these references, publications, and patents are incorporated herein by reference in their entirety.
Examples
The following examples are intended to illustrate certain preferred embodiments of the invention and are not intended to limit the scope of the invention. The following examples, specifically examples 1-8, demonstrate that compounds representative of the class of compounds described herein have been synthesized. Examples 9-17 show that compounds of formula (I) as synthesized in example 1 and compounds of formula (IIB) herein referred to as TTL1-TTL4 synthesized according to the method of example 1 and, in particular, examples 2-5, exhibit similar viability to untreated controls after treatment with increasing doses of up to 10 μ g/ml of compounds of formula (I) and compounds of formula (IIB) synthesized according to the method of example 1 and, in particular, examples 2-5, in mammalian cells presenting an acceptable preliminary model for human efficacy and safety, meaning that the inhibitory effect of the compounds evaluated on TNF- α synthesis is not mediated by direct cytotoxic effects.
Subsequent experiments with some preferred compounds OF THE invention demonstrated that TTL1 exhibited about ten (10) times stronger activity in inhibiting TNF- α and IL-1 synthesis than THE compound OF FORMULA (I) (THE SYNTHETIC COMPOUND OF FORMULA (I)). TTL3 containing an additional chemical modification showed about 100-fold greater activity than TTL 1. It is noted that, similar to the compounds of formula (I), neither TTL1 nor TTL3 significantly inhibited IL-6 synthesis.
Example 1
Stereoselective synthesis of compounds of formula (I) and (II)
The first stereoselective synthesis of the compound of formula (I) has been completed. Our synthesis project was initiated with (-) Wieland-Miesher ketone (107), see fig. 18, and referred to as the diels-alder cycloaddition reaction to construct the C ring of 101. The synthesis confirmed the proposed stereochemistry of 101 and represents an effective entry into the undeveloped biologically active diterpenes.
Root bark of Acanthopanax koreanum Nakai (Araliaceae), a shrub growing in Korea and fallen leaves every year, has been traditionally used as a tonic and sedative agent, and a drug for treating rheumatism and diabetes (Medicinal Plants of East and south Asia, Perry, L.M.; Metzger, J.Eds.; MIT Press, Cambridge, MA and London, 1980). During the pharmacological extract studies of this folk drug, Chung and co-workers have isolated a new diterpene and determined its structure, after which the diterpene was named Sankawa, U.J. Nat.Prod.1988, 51, 1080-1083; (b) Kang, H.S.; Kim, Y.H.; Lee, C.S.; Lee, J.J.; Choi.I.; Pyun, K.H.Cellular. Immunol.1996, 170, 212-221; (c) Kang, H.S.; Song, H.K.; Lee, J.J.; Pyun, K.H.H.; Choi, I.toraniatom.1998, Medfold, 7).
From a biosynthetic point of view, 101 belongs to a considerable family of pimaric diene diterpenes, which is best represented by pimaric acid (102) (Ruzicka, L.; Sternbach, L.; J.Am.chem.Soc.1948, 70, 2081-2085; Ireland, R.E.; Schiess, P.W.tetrahedron Lett.1960, 25, 37-43; Wenkert, E.; Buckwalter, B.L.J.Am.Chem.Soc.1972, 94, 4367-4372; Wenkert, E.; Chamberlin, J.W.J.Am.Soc.1959, 81, 688-693). The compounds of formula (I) are structurally characterized by a unique linkage via a rigid tricyclic core that may be critical to their pharmacological properties. In fact, the recent isolation of this compound has enabled the study of its biological activity and the examination of its pharmaceutical potential (Kang, H. -S.; Kim, Y. -H.; Lee, C. -S.; Lee, J. -J.; Choi., I.; Pyun, K. -H.Cellular Immunol.1996, 170, 212-221; Kang, H. -S.; Song, H.K.; Lee, J. -J.; Pyun, K. -H.; Choi, I.Mediators Inflamm 1998, 7, 257-259)). More specifically, it was found that antanthoic acid exhibits promising anti-inflammatory and anti-fibrotic activities presumably due to the inhibition of the pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-I). See, turbo cross fans, the molecular and hair Emerging roll in medicine, B. Beutler, ed.; raven Press, N.Y.1992; aggarwal, b.; purify, R. human cytokines. Their Role in Disease and Therapy; blackwell Science, inc: u.s.a., 1995; thorpe, r.; Mire-Sluis, a. cytokines; academic Press: san Diego, 1998; kurzrock, r.; talpaz, m.cytokines: interleukins and Their Receptors; kluwer Academic Publishers: u.s.a., 1995; szekanecz, z.; kosh, a.e.; kunkel, s.l.; striiter, r.m. clinical pharmicol.1998, 12, 377-390; camussi, g.; lupin, e.drugs 1998, 55, 613-620; newton, r.c.; decicco, C.P.J.Med.chem.1999, 42, 2295-2314.
The inhibition is concentration dependent and cytokine specific, because IL-6 or IFN-gamma (interferon-gamma) production is not affected under the same conditions. Furthermore, it was found that antathoic acid is active when administered orally and shows minimal toxicity in experiments with mice and rats.
This unusual structure, together with the promising pharmacological activities shown by 101, suggest that we extend synthetic research to this class of biologically important metabolites, see Xiang, a.x.; watson, d.a.; ling.t.; theodorakis, E.A.J.org.chem.1998, 63, 6774-6775; ling, t.; xiang, a.x.; theodorakis, E.A.Angew.chem.int.Ed.Engl.1999, 38, 3089-3091. This example provides a stereoselective total synthesis of (-) antanthoic acid and a compound of formula (II) as shown in examples 2-6, providing the basis for performing the total synthesis of a compound of formula (IIB). This example also identifies the structure and absolute stereochemistry of 101.
The strategy for the inverse synthesis of the antigenic acid is shown in FIG. 20. The C-ring of 101 is envisioned to be constructed by diels-alder cycloaddition, suggesting that we employ dienophiles 103 and appropriately substituted dienes such as 104 as the ideal coupling partners. See, Oppolzer, Comprehensive org, synthesis, Trost, b.m.ed. of W; oxford, n.y.; PergamonPress, 1991, 315-399. This reaction introduces unsaturation at the C9-C11 bond, and the desired stereochemistry at the C8 and C13 carbons, allowing a convenient fulcrum between the synthesis of the compound of formula (II) and the compound of formula (IIB). The dienes 104 can be prepared by functionalizing the ketones 105, wherein the C4 tetrasubstituted center of 105 is formed by stereocontrolled alkylation of the β -ketoesters 107. This analysis suggested the use of (-) Wieland-Miesher ketone 107 as the putative starting material. The use of such a scheme for synthesizing an aconthoic acid is depicted in FIGS. 21 and 23 as synthesis schemes 5 and 6. All compounds showed satisfactory spectral and analytical data.
Starting with optically pure enone 107, enone 107 was readily prepared via D-proline mediated asymmetric Robinson cyclization reaction (yield 75-80%, > 95% ee). See Buchschacher, p.; fuerst, a.; gutzwiller, J.org.Synth, Coll.Vol.VII 1990, 368-3372.). The C9 keto group of 107 was selectively ketalized and then reductively alkylated via an ketene functionality using methyl cyanoformate to afford the ketoester 106 in 50% overall yield. See Crabtree, s.r.; mander, L.N.; sethi, P.S.org.Synth.1992, 70, 256-263. To introduce the desired functionality at C4, a second reductive alkylation was performed, see Coates, r.m.; shaw, j.e.j.org.chem.1970, 35, 2597-2601; coates, r.m.; shaw, j.e.j.org.chem.1970, 35, 2601-2605. Compound 106 was first converted to the corresponding methoxymethyl ether 108 and ether 108 was treated with lithium in liquid ammonia and methyl iodide to obtain ester 110 as a single diastereomer in 58% overall yield. See Welch, s.c.; hagan, c.p.synthetic comm.1973, 3, 29-32; welch, s.c.; hagan, c.p.; kim, j.h.; chu, p.s.j.org.chem.1977, 42, 2879-2887; welch, s.c.; hagen.c.p.; white, d.h.; fleming, w.p.; trotter, j.w.j.am.chem.soc.1977, 99, 549-556. The stereoselectivity of this addition is due to the very preferential alkylation of the intermediate enol 109 on the side of the flattening-out with very little steric hindrance.
The C-ring is constructed using an existing bicyclic nucleus. The C-ring is formed by Diels-Alder reaction between methacrolein 103 (see, e.g., FIG. 21) and a sulfur-containing diene 104. 104 was initiated by acid catalyzed deprotection of the 110C 9 ketal, followed by alkylation of the resulting ketone 105 with a lithium acetylide-ethylenediamine complex. See Das, j.; dickinson, r.a.; kakushima, m.; kingston, g.m.; reid, g.r.; sato, y.; valenta, z.can.j.chem.1984, 62, 1103-1111). This procedure gives alkyne 111 in an overall yield of 86%, as an 8: 1 diastereomeric mixture at C9 (the isomers shown are predominant). At this point the diastereoselectivity of the Diels-Alder reaction was evaluated, along with the overall feasibility of using unfunctionalized dienes such as 112. For this purpose, the diastereomeric mixture of propargyl alcohol 111 is partially reduced (H)2Lindele catalyst) and dehydration (BF)3·Et2O), diene 112 was produced in 90% yield (coisone, j. -m.; pecher, j.; declercq, j. -p.; germanin.g.; van Meersche, M.Bull.Soc.Chim.Belg.1980, 89, 551-557). Diels-Alder cycloaddition of 112 with methacrolein (103) under neat conditions at 25 ℃ gave in quantitative yield a mixture of two diastereomeric aldehydes, which were reduced with sodium borohydride, thenAnd (5) separating. The resulting alcohols 114 and 115 were converted to the corresponding p-bromobenzoates (compounds 116 and 117, respectively) which were recrystallized from dichloromethane/ethanol to obtain crystals suitable for X-ray analysis (fig. 22).
The results of the X-ray analysis showed that the tricyclic system had the expected stereochemistry at C4 and confirmed that the Diels-Alder reaction was not internally directed. It is shown that methacrolein, when reacted with cyclopentadiene, produces the exo diels-alder product: kobuke, y.; fueno, t.; furukawa, J.J.am.chem.Soc.1970, 92, 6548-6553. This surprising observation can be reasonably explained by the steric repulsion exhibited by methyl groups: yoon, t.; danishefsky, s.j.; de Gala, S.Angew.chem.Int.Ed.Engl.1994, 33, 853-855. Further, after reduction, the main product of the cycloaddition is shown to be the alcohol 114 with the desired stereochemistry at the center of C8, thus confirming that the diene 112 reacts very preferentially from the α -face (bottom-side attack) with 103, see, e.g., fig. 21. Furthermore, these data indicate that the synthesis of the acontac acid requires an initial reversal of the orientation of the dienophile.
As discussed in examples 2-8 below, entirely new compounds of formula (IIB) were synthesized without inversion of the starting dienophile. By selecting appropriately substituted dienophiles, R of the compounds of the formula (IIB) can be selected essentially indefinitely11And R12A group.
Inversion of the dienophiles required for the synthesis of compounds of formula (I), their natural analogs, and compounds of formulae (II) and (IIA) is achieved by changing the orbital coefficient of the atoms at the ends of the dienes, which provides evidence for the use of heteroatom-containing dienes such as 104 during cycloaddition. See overlaman, l.e.; petty, c.b.; ban, t.; huang, G.T.J.am.chem.Soc.1983, 105, 6335-6338; trost, b.m.; ippen, j.; vladuchick, w.c.j.am.chem.soc.1977, 99, 8116-8118; cohen, t.; kozarych, Z.J.org.chem.1982, 47, 4008-4010; hopkins, p.b.; fuchs, p.l.j.org.chem.1978, 43, 1208-1217; petrzilka, m.; grayson, j.i. synthesis, 1981, 753-786). The construction of diene 104 and its use in synthesis 101 is shown in FIG. 23, FIG. 6.
Compound 104 was prepared by: the thiophenol group is subjected to a free radical addition to the alkyne 111 (greenras, c.w.; Hughman, j.a.; Parsons, p.j.chem.soc.chem.commun.1985, 889-890), and the allyl alcohol obtained is then subjected to POCl3Catalytic dehydration (Trost, B.M.; Jungheim, L.N.J.Am.chem.Soc.1980, 102, 7910-7925; Mehta, G.; Murthy, A.N.; Reddy, D.S.; Reddy, A.V.J.Am.chem.Soc.1986, 108, 3443-3452) (2 steps, 70% yield). Interestingly, attempts were also made to use BF3·Et2O undergoes dehydration, but the results prove to be ineffective in this case. Using a large number of available 104, we studied Diels-Alder reactions using 103 as a dienophile. Several heats (-78-80 ℃) and Lewis acids (BF) were tested3·Et2O，TiCl4，AlCl3And SnCl4) Catalytic Diels-Alder conditions. Using SnCl in dichloromethane at-20 deg.C4The best results were obtained with a 4.2: 1 diastereoisomeric mixture of aldehyde 118 in 84% yield. To simplify the characterization of the product and to enable sufficient separation, NaBH was used4The mixture is reduced and desulfurized using Raney nickel reduction. Thus, alcohols 119 and 120 were obtained in 91% overall yield. The structures of these compounds were determined by comparing the products isolated from the reactions of 103 and 112. The main diastereomer 120 was treated with Dess-Martin periodinane followed by Wittig methylation to introduce an olefinic functionality at the center of C13 to give 121 in 86% overall yield. The C-19 carboxylic acid is then deprotected. 121 was exposed to LiBr in refluxing DMF via SN2Displacement of the-acyloxy functional group gave the acontaic acid 101 in 93% yield. See Bennet, c.r.; cambie, r.c. tetrahedron 1967, 23, 927-941. Synthetic 101 has the same spectral and analytical data as the reported natural species.
This example provides a elegant, stereoselective method of synthesizing compound 101. The synthetic strategy is remarkable in that Diels-Alder reaction with diene 104 with methacrolein (103) generates stereochemistry at the C13 and C8 carbon centers. The process described for the synthesis of 101 requires 14 steps (starting from enone 107) with an overall yield of about 9%. The overall high efficiency and versatility of this strategy lays the foundation for our preparation of analogs designed to have improved pharmacological properties.
Examples 2 to 8
Stereoselective synthesis of compounds of formula (IIB) the process outlined in example 1 and depicted in FIG. 23, scheme 6, may be modified or shortened to prepare compounds of formula (II) or formula (IIB).
Example 2
Compound 114, referred to herein as TTL4, was synthesized by the method of example 1 as described in figure 21, which is referred to herein as TTL 4.
Example 3
The compound referred to herein as TTL2 was synthesized: compound 114 was synthesized by the method of example 1 as depicted in figure 21. Compound 114 was then reacted with 3.0 equivalents of LiBr in DMF at 160 ℃ for about 3 hours in a similar manner to the reaction described in figure 23, step (h), to give the compound referred to herein as TTL2 in about 93% yield.
Example 4
Compound 13, herein designated TTL3, was obtained by synthesis of a compound designated TTL3 as depicted in figure 14.
Example 5
Compound 13 was obtained by synthesis of a compound referred to herein as TTL1 as described in figure 14. This compound is referred to herein as TTL 1.
Example 6
Wherein R is15Is hydrogen, and R9And R15Independently selected from C1-C6Alkyl, and C1-C6Substituted alkyl compounds of formula (IIB) were synthesized by the method of example 1, except that in this example the dienophile was selected from those in which R is15Is hydrogen, and R9And R15Independently selected from C1-C6Alkyl, and C1-C6Substituted alkyl compounds of formula (III).
Example 7
In particular, wherein R14Is hydrogen, and R9And R15Independently selected from C1-C6Alkyl, and C1-C6Substituted alkyl compounds of formula (IIB) were synthesized by the method of example 1, except that in this example the dienophile was selected from those in which R is14Is hydrogen, and R9And R15Independently selected from C1-C6Alkyl, and C1-C6Substituted alkyl compounds of formula (III).
Example 8
Wherein R is14Is hydrogen, and R9And R15Independently selected from C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6Aryl compounds of formula (IIB) were synthesized by the method of example 1, except that in this example the dienophile was selected from those in which R is14Is hydrogen, and R9And R15Independently selected from C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6Aryl group of the formula (III).
Examples 9-17 materials and methods.
Mouse macrophage RAW 264.7(1 × 10)6/ml) week of different doses of the synthetic compound of formula (I), the synthetic compound of formula (I) and a group of analogues (diluted in 0.5% DMSO) for 30-60 minutes, followed by stimulation with different agents such as Lipopolysaccharide (LPS) or gram positive agents such as heat killed staphylococcus aureus (Staph aureus SAC). The levels of TNF-a, IL-I, IL-6, IL-10, IL-18 and other cytokines were determined in supernatants collected over a 72 hour period by enzyme-linked immunosorbent (elisa) or bioanalytical methods. Additional experiments were also conducted to evaluate the effect of the synthetic compounds of formulae (I), (II), (IIA) and (IIB) on specific cytokine signaling pathways such as Caspase activity (Nr-1, Nr.3), NF-. kappa. B, MAP-kinase activity (P38, ERK and JNK). Results
Preclinical experiments demonstrated that RAW 264.7 cells treated with increasing doses of compounds of formulae (I) and (IIB) at concentrations up to 10ug/ml, specifically the compounds referred to herein as TTL1 and TTL3, exhibited comparable viability to untreated controls, meaning that the inhibitory effect of the compounds of formulae (I) and (IIB) on TNF- α synthesis was not mediated by direct cytotoxic effects.
Subsequent experiments with the compound of formula (I) synthesized according to example 1, TTL1 (synthesized according to example 2) and TTL3 (synthesized according to example 4) demonstrated that TTL1 exhibited about 10-fold greater activity in inhibiting TNF- α and IL-1 synthesis than the compound of formula (I) synthesized according to example 1. TTL3 synthesized according to example 4, which contained an additional chemical modification, showed about 100-fold greater activity than TTL1 synthesized according to example 2. It is noted that, similar to the compound of formula (I) synthesized according to example 1, neither the analogs TTL1 nor TTL3 significantly inhibited IL-6 synthesis. TTL1 exhibited ten (10) times greater activity than the compound of formula (I) synthesized according to example 1 in inhibiting TNF- α and IL-1 synthesis.
TTL3 containing an additional chemical modification showed about 100-fold greater activity than TTL 1. It is also noted that, similar to the compound of formula (I) synthesized according to example 1, neither analog significantly inhibits IL-6 synthesis.
TABLE 1
Compounds of formula (I) and TTL1 inhibit LPS-induced TNF-alpha synthesis
| LPS | Formula (I) (0.1. mu.g/ml) | Formula (I) (1. mu.g/ml) | Formula (I) (10. mu.g/ml) | TTL1(0.1μg/ml) | TTL1(1μg/ml) | TTL1(5.4μg/ml) | |
| TNF-α(ng/ml) | 120 | 108 | 67 | 50 | 57 | 60 | 38 |
TABLE 2
Compounds of formula (I) and TTL1 inhibit SAC-induced TNF-alpha synthesis
| SAC | Formula (I) (0.1. mu.g/ml) | Formula (I) (1. mu.g/ml) | Formula (I) (10. mu.g/ml) | TTL1(0.1μg/ml) | TTL1(1μg/ml) | TTL1(5.4μg/ml) | |
| TNF-α(ng/ml) | 385 | 410 | 275 | 165 | 250 | 285 | 150 |
TABLE 3
Compounds of formula (I) and TTL1 inhibit SAC-induced IL-1 synthesis
| SAC | Formula (I) (0.1. mu.g/ml) | Formula (I) (1. mu.g/ml) | Formula (I) (10. mu.g/ml) | TTL1(0.1μg/ml) | TTL1(1μg/ml) | TTL1(5.4μg/ml) | |
| IL-1α(pg/ml) | 700 | 1350 | 1050 | 350 | 950 | 400 | 300 |
TABLE 4
The compound of formula (I) and TTL1 do not inhibit SAC-induced IL-6 synthesis
| SAC | Formula (I) (0.1. mu.g/ml) | Formula (I) (1. mu.g/ml) | Formula (I) (10. mu.g/ml) | TTL1(0.1μg/ml) | TTL1(1μg/ml) | TTL1(5.4μg/ml) | |
| IL-6(ng/ml) | 75 | 65 | 90 | 80 | 83 | 86 | 65 |
TABLE 5
TTL3 inhibits SAC-induced TNT-alpha synthesis
| Is not stimulated | SAC | 0.001μg/ml | 0.01μg/ml | 0.1μg/ml | 1μg/ml | 10μg/ml | |
| TNF-α(ng/ml) | 5 | 375 | 80 | 75 | 85 | 60 | 80 |
TABLE 6
TTL3 inhibits SAC-induced IL-1 synthesis
| Is not stimulated | SAC | 0.001μg/ml | 0.01μg/ml | 0.1μg/ml | 1μg/ml | 10μg/ml | |
| IL-1α(pg/ml) | 0 | 650 | 200 | 220 | 190 | 180 | 170 |
TABLE 7
TTL3 inhibits LPS-induced TNF-alpha synthesis (TNF-alpha (ng/ml))
| LPS(1μg/ml)+TTL3(μg/ml) | ||||||
| LPS alone | (1×10-7) | (1×10-6) | (1×10-5) | (1×10-4) | (1.0) | (1×102) |
| 88 | 41 | 18 | 10 | 15 | 13 | 4 |
TABLE 8
TTL3 inhibits mortality following administration of LPS/D-Gal
*All treatments were given intraperitoneally, with TTL3 given 45 minutes prior to LPS administration
| Treatment of* | Mortality rate 24 hours | Death rate of 48 hours |
| LPS/D-Gal | 10/10 | 10/10 |
| LPS/D-Gal+DMSO | 8/10 | 9/10 |
| LPS/D-Gal+TTL3 | 2/10 | 2/10 |
While particular embodiments of the present invention have been shown and described in detail to illustrate the application and principles of the invention, it will be understood that the invention may be embodied otherwise without departing from the principles of the invention.
Claims (40)
1. A compound having the following chemical structure:wherein:
R1selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12An aryl group;
R2and R9Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Alcohol, C1-C12Acyl, and C5-C12An aryl group;
R3-R5、R7、R8and R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12An aryl group;
R6selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12An alkynyl group;
R10selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12An aryl group; and is
R14And R15Independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6An aryl group;
wherein said compound comprises a prodrug ester of the above compound, and acid addition salts thereof.
2. The compound of claim 1, wherein:
R1selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohol, (C)1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group.
3. The compound of claim 1, wherein:
R1selected from hydrogen, halogen, COOH, C1-C6Carboxylic acid, C1-C6Acyl halide, C1-C6Acyl radical, C1-C6Esters, C1-C6Secondary amide, (C)1-C6)(C1-C6) Tertiary amides, C1-C6Alcohol, (C)1-C6)(C1-C6) Ether, C1-C6Alkyl radical, C1-C16Substituted alkyl, C2-C6Alkenyl radical, C2-C12Substituted alkenyl, and C5-C6And (4) an aryl group.
4. The compound of claim 1, wherein R1Is selected from C2-C6Esters and C1-C6An acyl group.
5. The compound of claim 1, wherein R1Is selected from C2-C6And (3) an ester.
6. The compound of claim 1, wherein R10Is selected from C2-C6Alkyl and C2-C6An alkenyl group.
7. The compound of claim 1, wherein R3-R5、R7、R8、R11-R15Each is hydrogen.
8. The compound of claim 7, wherein R3-R5、R7、R8、R11-R15Each is hydrogen; r2、R6And R9Each is methyl; and R is10Is CH2。
9. The compound of claim 1, wherein R15Is hydrogen, and R14Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohol, (C)1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group.
10. The compound of claim 1, wherein R15Is hydrogen, and R14Selected from hydrogen, halogen, C2-C6Alcohol, C2-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, and C5-C6And (4) an aryl group.
11. The compound of claim 1 wherein said compound is selected from the group consisting of TTL1, TTL2, TTL3, and TTL4 and prodrug esters and acid addition salts thereof.
12. The compound of claim 1 wherein said compound is selected from the group consisting of TLL1 and TTL3 and their prodrug esters and their acid addition salts.
13. A method of treating a disease selected from the group consisting of inflammation, tuberculous pleurisy, rheumatoid pleurisy, cancer, cardiovascular disease, redness of the skin, diabetes, transplant rejection, otitis media (inner ear infection), sinusitis, and viral infection, comprising:
contacting a compound with living tissue of said animal, wherein said compound is a compound of claim 1.
14. The method of claim 13, wherein the compound is a compound of claim 2, 3, 4, 5,6, 7,8, 9, 10, 11, or 12.
15. A compound having the following chemical structure:wherein:
if any R3-R5、R7、R8、R11-R15Not being hydrogen, R2Or R6Or R9Not being methyl, or R10Is not CH2Then R is1Selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12An aryl group; however, it is possible to use a single-layer,
if all R are3-R5、R7、R8、R11-R13Are all hydrogen, R2、R6And R9Are each methyl, and R10Is CH2Then R is1Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohols, (C) other than methylacetyl ether1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12An aryl group;
R2and R9Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, C1-C12Alcohol, C1-C12Acyl, and C5-C12An aryl group;
R3-R5、R7、R8and R11-R13Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12An aryl group;
R6selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12An alkynyl group;
R10selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12An aryl group; and is
R14And R15Independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C6Alcohol, and C5-C6An aryl group;
wherein said compound comprises a prodrug ester of the above compound, and acid addition salts thereof.
16. The compound of claim 15, wherein:
R1selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohols, (C) other than methylacetyl ether1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12And (4) an aryl group.
17. The compound of claim 15, wherein:
R1selected from hydrogen, halogen, COOH, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C1-C12Esters, C1-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C1-C12Alcohol, (C)1-C12)(C1-C12) Ether, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group.
18. The compound of claim 25, wherein R1Is selected from C2-C12Esters and C1-C12An acyl group.
19. The compound of claim 15, wherein R1Is selected from C2-C6And (3) an ester.
20. The compound of claim 25, wherein R10Is selected from C2-C6Alkyl and C2-C6An alkenyl group.
21. The compound of claim 15, wherein R3-R5、R7、R8、R11-R15Respectively hydrogen.
22. The compound of claim 21, wherein R3-R5、R7、R8、R11-R15Are each hydrogen; r2、R6And R9Are each methyl; and R is10Is CH2。
23. The compound of claim 15, wherein R15Is hydrogen, and R14Selected from hydrogen, halogen, C1-C12Carboxylic acid, C1-C12Acyl halide, C1-C12Acyl radical, C2-C12Esters, C2-C12Secondary amide, (C)1-C12)(C1-C12) Tertiary amides, C2-C12Alcohol, (C)1-C12)(C1-C12) Ether, C2-C12Alkyl radical, C1-C12Substituted byAlkyl radical, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C5-C12And (4) an aryl group.
24. The compound of claim 15, wherein R15Is hydrogen, and R14Selected from hydrogen, halogen, C2-C6Alcohol, C2-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, and C5-C6And (4) an aryl group.
25. A method of treating a disease selected from the group consisting of inflammation, tuberculous pleurisy, rheumatoid pleurisy, cancer, cardiovascular disease, redness of the skin, diabetes, transplant rejection, otitis media (inner ear infection), sinusitis, and viral infection, comprising:
identifying an animal having the disease; and contacting a compound with living tissue of the animal, wherein the compound is a compound of claim 15.
26. The method of claim 25, wherein the compound is a compound of claim 16, 17, 18, 19, 20, 21, 22, 23, or 24.
27. A method of treating a disease selected from the group consisting of tuberculous pleurisy, rheumatoid pleurisy, cancer, cardiovascular disease, redness of the skin, diabetes, transplant rejection, otitis media (inner ear infection), sinusitis, and viral infection, comprising:
identifying an animal having the disease; and contacting a compound with living tissue of said animal, wherein said compound is selected from the group consisting of (a) antathoic acid, (b) (-) -pimpinel-9 (11), 15-dien-19-ol, (c) (-) -pimpinel-9 (11), 15-dien-19-oic acid, (d) (-) -pimpinel-9 (11), 15-dien-19-ol 19-acetate, (e) (-) -pimpinel-9 (11), 15-diene, and (f) a methyl ester analog of antathoic acid.
28. The method of claim 27, wherein said compound is an anticathoic acid.
29. The method of claim 27, wherein the compound is (-) -pimara-9 (11), 15-dien-19-ol.
30. The method of claim 27, wherein the compound is (-) -pimara-9 (11), 15-diene-19-acid.
31. The method of claim 27, wherein the compound is (-) -pimara-9 (11), 15-dien-19-ol 19-acetate.
32. The method of claim 27, wherein the compound is (-) -pimara-9 (11), 15-diene.
33. The method of claim 27, wherein said compound is a methyl ester analog of acanthoic acid.
34. A compound having the following chemical structure:
wherein R is1And R1’Are covalently linked, and wherein
R1And R1’Are each selected from C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl, and C2-C6A substituted alkenyl group;
R2and R9And R2’And R9’Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C1-C12Acyl group and C5-C12An aryl group;
R3-R5、R7、R8and R11-R15And R3’-R5’、R7’、R8’And R11’-R15’Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12An aryl group;
R6and R6’Independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12An alkynyl group; and is
R10And R10’Independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group.
35. A compound having the following chemical structure:
wherein R is1And R1’Are covalently linked, and wherein
R1And R1’Are each selected from C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl, and C2-C6A substituted alkenyl group;
R2and R9And R2’And R9’Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C1-C12Acyl group and C5-C12An aryl group;
R3-R5、R7、R8and R11-R15And R3’-R5’、R7’、R8’And R11’-R15’Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12An aryl group;
R6and R6’Independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12An alkynyl group; and is
R10And R10’Independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group.
36. A compound having the following chemical structure:
wherein R is1And R1’Are covalently linked, and wherein
R1And R1’Each independently selected from C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl, and C2-C6A substituted alkenyl group;
R2and R9And R2’And R9’Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted byAlkenyl radical, C2-C12Alkynyl, and C1-C12Acyl group and C5-C12An aryl group;
R3-R5、R7、R8and R11-R15And R3’-R5’、R7’、R8’And R11’-R15’Each independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, C2-C12Alkynyl, and C5-C12An aryl group;
R6and R6’Independently selected from hydrogen, halogen, C1-C12Alkyl radical, C1-C12Substituted alkyl, C2-C12Alkenyl radical, C2-C12Substituted alkenyl, and C2-C12An alkynyl group; and is
R10And R10’Independently selected from hydrogen, halogen, CH2、C1-C6Alkyl radical, C1-C6Substituted alkyl, C2-C6Alkenyl radical, C2-C6Substituted alkenyl, C1-C12Alcohol, and C5-C12And (4) an aryl group.
37. The compound of claim 34, 35 or 36, wherein
R1And R1Together constitute the following structure:
-CH2-O-CO-(CH2)n-CO-O-CH2-，
wherein n is a positive integer no greater than 8.
38. A method of treating a disease selected from the group consisting of tuberculous pleurisy, rheumatoid pleurisy, cancer, cardiovascular disease, redness of the skin, diabetes, transplant rejection, otitis media (inner ear infection), sinusitis, and viral infection comprising contacting a compound with living tissue of the animal, wherein the compound is a compound of claim 34, 35, or 36.
39. A synthetic method for preparing a compound of claims 1-10 or 11, comprising the steps of:
subjecting a diene having two or more rings to a Diels-Alder reaction with a dienophile compound to produce a compound having 3 or more rings; and
preparing a compound according to claim 1-10 or 11.
40. A synthetic method for preparing a compound of claim 15-23 or 24, comprising the steps of:
subjecting a diene having two or more rings to a Diels-Alder reaction with a dienophile compound to produce a compound having 3 or more rings; and
preparing a compound according to claim 15-24 or 25.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US60/134,295 | 1999-05-14 | ||
| US60/186,853 | 2000-03-03 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HK1048110A true HK1048110A (en) | 2003-03-21 |
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