(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY(PCT)
`
`(19) World Intellectual Property Organization
`International Bureau
`
`(43) International Publication Date
`14 June 2001 (14.06.2001)
`
`UE
`
`PCT
`
`(10) International Publication Number
`WO 01/42231 A2
`
`Columbia V7R 4P3 (CA). LI, Jianxiong [CA/CA]; 117
`Buckingham Drive, Port Moody, British Columbia V3H
`274 (CA). HU, Kaiji
`[CA/CA]; 6940 Curtis Street,
`Burnaby, British Columbia VSB 2B1 (CA). ZHU, Jiang
`
`{21) International Application Number:=PCT/CA00/01433 [CA/CA]; 406 Louis Riel House, Burnaby, British Colum-
`bia V5A 186 (CA).
`
`CO7D 303/14,
`(51) International Patent Classification’:
`303/24, 303/40, A61K 31/355, 31/05, 31/09, A61P 17/06,
`29/00
`
`(22) International Filing Date: 6 December 2000 (06.12.2000)
`
`(25) Filing Language:
`
`(26) Publication Language:
`
`English
`
`English
`
`(30) Priority Data:
`60/168,758
`60/173,300
`
`6 December 1999 (06.12.1999) US
`28 December 1999 (28.12.1999)
`US
`
`(74) Agent: BLAKE, CASSELS & GRAYDON LLP; 20th
`Floor, 45 O’ConnorStreet, Ottawa, Ontario K1P 1A4 (CA).
`
`(81) Designated States (national): AE, AG, AL, AM, AT, AU,
`AZ, BA, BB, BG,BR, BY, BZ, CA, CH, CN, CR, CU, CZ,
`DE, DK, DM,DZ,EE,ES, FI, GB, GD, GE, GH, GM, HR,
`HU, ID,IL,IN,IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR,
`LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ,
`NO, NZ, PL, PT, RO, RU, SD,SE, SG, SI, SK, SL, TJ. TM,
`TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW.
`
`(84) Designated States (regional): ARIPO patent (GH, GM,
`KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZW), Eurasian
`patent (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), European
`patent (AT, BE, CH, CY, DE, DK,ES, FI, FR, GB, GR,IE,
`IT, LU, MC,NL,PT, SE, TR), OAPIpatent (BF, BJ, CF,
`CG,CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG).
`
`(71) Applicant (for all designated States except CN, US):
`WELICHEM BIOTECHINC.[CA/CA];Suite 13, 3871
`North Fraser Way, Bumaby, British Columbia V5] 5G6
`(CA).
`
`(71) Applicant and
`(72) Inventor (for CN, US only): CHEN, Genhui [CA/CA];
`725 Louis Riel, Burnaby, British Columbia V5A 186 (CA).
`
` LMUARTYAMAA
`
`Published:
`(72) Inventors; and
`(75) Inventors/Applicants (for US only): WEBSTER, John, — Without international search report and to be republished
`M. [CA/CA}; 551 Molina Road, North Vancouver, British
`upon receipt ofthat report.
`.
`{Continued on next page]
`
`(54) Title: ANTI-INFLAMMATORY AND PSORIASIS TREATMENT AND PROTEIN KINASE INHIBITION BY HYDROX-
`YLSTILBENES AND NOVEL STILBENE DERIVATIVES AND ANALOGUES
`
`Effect of 3,5-dihydroxy-4-isopropylistilbene on
`crystal induced neutrophil activation
`
`600
`g
`500
`9
`400
`a
`a % 300
`52
`=
`«200
`g
`100
`Qo
`0
`
`
`
`H Control
`
`03,5-dihydroxy-4-
`i
`yistilbene,
`
`T
`
`1
`
`5
`
`7
`.
`Time (min)
`
`10
`
`a
`<
`=
`a
`Q
`—,
`—
`
`==
`
`(57) Abstract: Disclosed herein are compositions containing hydroxylstilbenes or their derivatives or analogues. The compositions
`are useful to inhibit protein kinease, and for the treatmentof inflammatory diseases, including psoriasis , multiple sclerosis, rhuma-
`= toid arthritis, restinosis, inflammatorybowel disease, and inflammatory lung disease. Theyare alsousefulto treat surgical adhesions
`and graft rejection. Novel derivatives and analogues are also disclosed.
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`[MAINTAINATTRAARTA
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`For two-letter codes and other abbreviations, refer to the "Guid-
`ance Notes on Codes andAbbreviations” appearing at the begin-
`ning ofeach regular issue ofthe PCT Gazette.
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`Anti-inflammatory and psoriasis treatment and protein kinase inhibition by
`
`hydroxylstilbenes and novel stilbene derivatives and analogues
`
`BACKGROUND OF THE INVENTION
`
`The stilbenes isolated from Photorhabdus species bacteria are known to have
`
`antibiotic activity. See V.J. Paul, S. Frautschv, W. Fenical, and K.H. Nealson,. Journal
`
`of Chemical Ecology, 7: 589-597 (1981), and K. Hu, J. Li, W. Wang, H. Wu, H. Lin
`
`and J. M. Webster, Canadian Journal of Microbiology. 44: 1072-1077 (1998).
`
`However, these compounds have not been shown to have otherbiological activity. A
`
`similar compound,resveratrol, has been disclosed as having cancer preventive (Jang et
`
`al. 1997) and protein kinase C inhibitory (Garcia-Garcia et. al., 1999) activities.
`
`There are many common conditions that cannot be treated successfully by antibiotics.
`
`Someofthese are inflammatory diseases. The compoundsof the invention possess
`
`specific anti-inflammatory properties.
`
`Inflammatory diseases, whether of a chronic or acute nature, represent a substantial
`
`problem in the healthcare industry. Chronic inflammation is considered to be
`
`inflammation of a prolonged duration (weeks or months) in which active inflammation,
`
`tissue destruction and attempts at healing are proceeding simultaneously (Robbins
`
`Pathological Basis ofDisease by R. S. Cotran, v. Kumar , and S. L. Robbins, W. B,
`
`Saunders Co., p. 75, 1989). Although chronic inflammation can follow an acute
`
`inflammatory episode, it can also begin as an insidious processthat progresses with
`
`time, for example, as a result of a persistent infection ( ¢.g. , tuberculosis, syphilis,
`
`fungal infection) which causes a delayed hypersensitivity reaction, prolonged exposure
`
`to endogenous(e.g., elevated plasmalipids) or exogenous(e.g., silica, asbestos,
`
`cigarette tar, surgical sutures) toxins, or, autoimmunereactions against the body's own
`
`tissues (e.g., rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis,
`
`psoriasis). Chronic inflammatory diseases therefore, include many common medical
`
`conditions such as rheumatoid arthritis, restenosis, psoriasis, multiple sclerosis, surgical
`
`adhesions, tuberculosis, and chronic inflammatory lung and airwaydiseases(e.g.,
`
`asthma, pneumoconiosis, chronic obstructive pulmonary disease, nasal polyps and
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`pulmonary fibrosis).
`
`Psoriasis is a common, chronic inflammatory skin disease. characterized by
`
`rapid multiplication and turnoverofthe epithelial cells with a consequent thickening of
`
`the epidermis, as well as inflamed swollen skin lesions covered with silvery white
`
`scaling patches and raised, inflamed, thickened and scaly lesions, which itch, burn,
`
`sting and bleed easily.It is therefore a disease characterized not only by inflammation,
`
`but also by proliferation ofcells. In the respect of proliferation, therefore, it has
`
`similarities to cancers, and both psoriasis and cancers can be described as proliferative
`
`diseases. In approximately 10% ofpatients, psoriasis is accompanied by pronounced
`
`arthropathic symptomsthat are similar to the changes seen in rheumatoidarthritis.
`
`Approximately 2 to 3% of the U.S. population suffers from psoriasis, with 250,000 new
`
`cases being diagnosed each year. The compoundsofthe invention possess specific
`
`activity against psoriasis.
`
`Antibiotic activity is also not a predictorof utility as a protein kineaseinhibitor.
`
`Protein kinease is implicated in many diseases, including cancers, as well as being a
`
`factor in diabetes, heromatous plaque and epidermalproliferation (including psoriasis.).
`
`The compoundsofthe invention possess specific protein kinase inhibiting activity.
`
`It is believed that protein kinase inhibitors may be of great importance in the
`
`control of uncontrolled cellular reproduction,i.e. in cellular reproduction disorders.
`
`For example DNA-PKisa serine/threonine protein kinase that is composed of a very
`
`large catalytic polypeptide and a DNA binding/targeting regulatory subunit (Ku
`
`autoantigen). Ku was first recognized as a heterodimeric (p70/p80) nuclear
`
`phosphoprote in that reacted with sera from patients suffering from autotmmune
`
`diseases lupus erythematosus and scleroderma polymyositis. Casein kinase II (Ck2)is a
`
`serine/threonine kinase that phosphorylates acidic protein such as casein. Ck2 has been
`
`shown to play multiple roles inside the cell and can be activated by numereous growth
`
`factors, hormones and cytokines. Ck2 has multiple substrate targets inside the cell
`
`which are untimately involved in the regulation of DNA, RNAandprotein synthesis.
`
`Ck2 plays a role in controlling mitogenic signalling, neuritogenesis. Ck2 has been
`shownto be involved with numerousdisease states. Elevated Ck2 levels have been
`
`demonstrated in solid human tumors and rapid proliferating non-neoplastic tisue such
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`3
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`as colorectal mucosa. Ck2 activity was much higher in metastatic melanomaand in
`
`cells transformed by human cytomegalovirus. Infection of animals with protoxoan
`
`parasite resulted in fatal lymphoproliferative syndromethat is asssociated with the over
`
`expression of Ck2. Ck2 activity has also been demonstrated to be elevated in
`Alzheimers disease.
`
`Amsonet al. (1989) showed that the 33-kD productof the PIM geneis highly
`
`expressed in the liver and spleen during fetal hematopoiesis. In contrast, it is only
`
`slightly expressed in circulating granulocytes in adults. It was over expressed in
`
`hematopoietic malignancies, particularly in myeloid and lymphoidacute leukemias. The
`
`results implicd a physiologic role of the Pim1 oncogene during hematopoietic
`
`development and a deregulation of the gene in various leukemias. Saris et al. (1991)
`
`provided evidence that both the murine and the human Pim1gene productsare protein-
`
`serine/threonine kinases. In the mouse,at any rate, they showedthat the gene encodes
`
`both a 44-and a 34-kD protein, the former being an amino-terminal extension of the
`
`latter which is synthesized by alternative translation initiation at an upstream CUG
`
`codon. Ark et al. (1991) provided refined mapping of the Pim-1 focus in the mouse and
`
`used the Pim-1 gene as a markerfor further genetic analysis of t-haplotypes on mouse
`
`chromosome 17. To understand the function of Pim-! andits role in hematopoietic
`
`development, Laird et al. (1993) generated mice deficient in Pim-1 function. Pim-1-
`
`deficient mice were ostensibly normal, healthy, and fertile; however, detailed analysis
`
`demonstrated a correlation of Pim-1 deficiency with erythrocyte microcytosis, whereas
`
`over expression of Pim-1 in transgenic mice resulted in erythrocyte macrocytosis.
`
`Recent studies on the molecular basis or neoplastic transformation have
`
`identified a family of genes, designated oncogenes, whose aberrant expression causes
`
`tumorigenesis. For example, the RNA tumourviruses possess such an oncogene
`
`sequence whose expression determines neoplastic conversion of infected ceils. The
`
`tyrosine kinase Lek are expressed primarily in different types of hematopoietic cells.
`
`The Lck protein is found in thymocytes and mature T cells and has been reported to be
`
`expressed in mature mousesplenic B cells. Campbell et al. (1992). Lek inhibitors are of
`
`value in the treatment of a numberof such disorders (for example, the treatment of
`
`autoimmunediseases), as Lck inhibition blocks T cell activation. The treatment of T
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`cell mediated diseases, including inhibition of T cell activation and proliferation, is a
`
`particularly preferred embodimentof the present invention. Compounds which
`
`selectively block T cell activation and proliferation are preferred.
`
`Accordingly, a specific inhibitor of these kinases can be usefulin investigating
`
`the mechanism of cancerogenesis, cell proliferation, differentiations and
`
`autoimmunology andit can be effective in prevention and chemotherapy of cancer and
`
`other pathological proliferative conditions. Hence the compoundsaccording to the
`
`present invention can be usefulin the treatment of pathological proliferation and
`
`autoimmunedisorders in mammals, including humans. A human or animal,e.g. a
`
`mammal, can thus be treated by a method comprising the administration thereto of a
`
`therapeutically effective amountof one of the compoundsof the invention.
`Amelioration of the disease state or disorder from which the human or animalis
`
`suffering can be achieved. Typical examples of such disorders are benign and
`
`malignant tumours, including leukaemia such as mycloblastic leukaemia, lymphoma,
`
`sarcoma, neuroblastoma, Wilm's tumour, malignant neoplasm ofthe bladder, breast,
`
`lung or thyroid, neoplasias of epithelialorigin, such as mammacarcinoma. Moreover,
`
`they can be useful in the treatment of epidermal hyper-proliferation, such aspsoriasis.
`
`The compoundsof the invention can also be useful in inhibiting the developmentofthe
`
`heromatous plaque and restenosis, in the control of angiogenesis, as anti-metastatic
`
`agents andin treating diabetic complications. They have also utility in the control of
`
`immunesystem diseases, ¢.g. as immuno-suppressants,as far as protein tyrosine
`kinases are involved in these diseases.
`
`Transplant rejection (graft v. host disease) and surgical adhesionsare also affected by
`
`protein kinases. The protein kinease inhibition and anti-inflamatory properties of the
`
`compoundsofthis inventiongiverise to utility in surgery to reduce transplantrejection
`
`and surgical adhesions.
`
`In a particular embodiment, the compoundsof the present inventionare useful for the
`
`treatment ofthe aforementioned exemplary disorders irrespective oftheir etiology, for
`
`example,for the treatment of transplant rejection, rbeumatoidarthritis, multiple
`
`sclerosis, chronic obstructive pulmonary disease, inflammatory bowel disease, lupus,
`
`graft vs host disease, T-cell mediated hypersensitivity disease, psoriasis, Hashimoto's
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`thyroiditis, Guillain-Barre syndrome, cancer, contact dermatitis,allergic disease such as
`
`allergic rhinitis, asthma, ischemic or reperfusion injury, or atopic dermatitis
`
`SUMMARYOF THE INVENTION
`
`The present invention relates to compositions containing the hydroxylated
`
`stilbenes and their derivatives and analogues which are useful in the treatment of
`
`inflammatory diseases, for the treatment of psoriasis, and for the inhibition of protein
`kineases.
`
`The invention also comprises the novel synthesis methods used to make the
`
`compounds. These methods render commercial manufacture possible and do not
`
`require recovery from insects.
`
`The present invention also comprises novel compounds,of the formula II given below.
`
`DETAILED DESCRIPTION OF THE INVENTION
`
`20
`
`25
`
`30
`
`In one preferred embodiment, this invention relates to compositions containing an
`
`antinflammatory amount of a compoundof the formula (FormualaI):
`OR,
`
`yof
`Xn =
`
`R
`
`OR2
`
`wherein
`
`R is selected from the group consisting of substituted and unsubstituted alkyl,
`
`substituted and unsubstituted alkenyl, substituted and unsubstituted cyclic alkyl,
`
`substituted and unsubstituted aryl, halide, alkoxyl, acyl, alkylsulfoxyl, alkylsulfonyl,
`
`alkylthiol, hydroxyl, thiol, substituted and unsubstituted amino groups,
`
`X is selected from the group consisting of substituted and unsubstituted alkyl,
`
`substituted and unsubstituted alkenyl, substituted and unsubstituted cyclic alkyl,
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`substituted and unsubstituted aryl, substituted and unsubstituted amino groups,nitro,
`
`carboxylic, acyloxyl such as acetoxyl, hydroxyl, alkoxyi, halide, acyl, alkylsulfoxyl,
`
`alkylsulfonyl, alkyithiol, aryl and heterocyclic ring,
`nis 0-5, and
`
`Ri and R2 are selected from the group of acyl, hydrogen and alkyl group,
`
`and the pharmaceutically acceptable salts thereof.
`
`10
`
`15
`
`The compoundsofthe invention are particularly useful against psoriasis, and, in a
`
`preferred embodiment, the compoundis present in an antipsoriatic amount.
`
`The compoundsof the invention are also useful as protein kinase inhibitors, and in a
`
`preferred embodiment, the compoundis present in an amountto inhibit protein kinase
`
`activity.
`
`The inventions also comprise methodsof treating inflammation,or psoroasis, or trating
`
`conditions whichare treatable by protein kinease inhibition, by administering a
`
`pharaceutically effective amount of the compoundsof the invention.
`
`20
`compounds whereinRis an alkyl group, R1 and R2 are hydrogen. Particularly
`
`Preferred compoundsfor use in the compositions of the present invention are those
`
`preferred are thosc compounds wherein R are branched short chain alkyl group, R1 and
`
`R2 are hydrogen. A highly preferred compoundis 3,5-dihydroxy-4-isopropylstilbene.
`
`25
`
`30
`
`In another preferred embodiment, R is selected from the group consisting of
`
`substituted and unsubstituted alkyl with carbon between I to 18, substituted and
`
`unsubstituted alkenyl with carbon between | to 18, substituted and unsubstituted cyclic
`
`alkyl with carbon between 1 to 18, halide, alkoxyl carbon between 1 to 18, acyl groups
`
`having a CO group and 1 to 17 additional carbon atoms,alkylsulfoxyl with with carbon
`
`between 1 to 18, alkylsulfonyl with carbon between 1 to 18, alkylthiol with carbon
`
`between 1 to 18, hydroxyl, thiol, X is selected from the group consisting of hydrogen,
`
`substituted and unsubstituted alkyl with carbon between 1 to 18, substituted and
`
`unsubstituted alkenyl with carbon between | to 18, substituted and unsubstituted cyclic
`
`alkyl with carbon between 1 to 18, substituted and unsubstituted amino groups,nitro,
`
`carboxylic, acyloxyl such as acetoxyl, hydroxyl, alkoxyl, halide, acyl group as C)-
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`Cy;CO- alkylsulfoxyl with carbon between | to 18, alkylsulfonyl with carbon between
`
`1 to 18, alkylthiol with carbon between | to 18, and substituted and unsubstituted
`
`phenyl
`
`nis 0-5,
`
`RI and R2 are selected from the group of acy! with carbon between | to 18, hydrogen
`
`and alkyl groups with carbon between | to 18,
`
`10
`
`and the pharmaccutically acceptable salts thereof.
`
`In a further preferred embodiment, R is selected from the group consisting of
`
`substituted and unsubstituted alkyl with carbon between | to 18, substituted and
`
`unsubstituted alkenyl with carbon between 1 to 18, substituted and unsubstituted cyclic
`
`alkyl with carbon between 1 to 18, and halide
`
`X is selected from the group consisting of methyl, acetoxyl, hydroxyl, methoxyl,
`
`halide, acetyl
`
`20
`
`nis 0-5, and
`
`R1 and R2 are selected from the group of acyl with carbon between 1 to 18, hydrogen
`
`and methyl groups,
`
`and the pharmaceutically acceptablesalts thereof.
`
`In a particularly preferred embodiment, the composition contains 3,5-dihydroxy-4-
`
`isopropylstilbene.
`
`This inventionalso relates to novel compounds and compositions containing an
`
`25
`
`30
`
`¢» °
`*e
`{ "
`
`anti- inflammatory amount of a compoundofthe formula II:
`OR
`
`OR?
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`wherein:
`
`R is selected from the group consisting of hydrogen, substituted and unsubstituted
`
`alkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted cyclic alkyl,
`
`substituted and unsubstituted aryl, halide, alkoxyl, acyl, alkylsulfoxyl, alkylsulfonyl,
`
`alkyithiol, hydroxyl, thiol, substituted and unsubstituted amino groups,
`
`X is selected from the group consisting of substituted and unsubstituted alkyl,
`
`substituted and unsubstituted alkenyl, substituted and unsubstituted cyclic alkyl,
`
`substituted and unsubstituted aryl, substituted and unsubstituted amino groups, nitro,
`
`carboxylic, acyloxyl such as acetoxyl, hydroxyl, alkoxyl, halide, acyl, alkylsulfoxyl,
`
`alkylsulfonyl, alkylthiol, aryl and heterocyclic ring
`
`nis 0-5, and
`
`RI and R2 are selected from the group of acyl, hydrogen and alkyl group.
`
`In addition to the free bases of formula I and II, the pharmaceutically acceptablesalts
`
`and the like are contemplated as being within the scope ofthis invention.
`
`20
`
`The compoundsofthis invention and the inventive compositions encompass both the
`
`trans and cis stereochemical forms of the compounds,and mixturesofthe trans and cis
`forms.
`
`25
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`Figure | is a graph showingthe effect of 3,5-dihydroxy-4-isopropylstilbene on crystal
`
`induced neutrophil activation.
`
`30
`
`Figure 2 is a graph showingthe effect of 3,5-dihydroxy-4-isopropylstilbene on fMLP
`
`induced neutrophil activation.
`
`Example 1 - Synthesis of the Inventive Compounds
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`The compoundsuseful in the compositions of the present invention may be prepared
`
`from 3,4-dihydroxybenzoic acid and 4-bromo-3,4-dihydroxybenzoic acid of the
`
`formula
`
`OOH
`
`COOH
`
`And
`
`HO
`
`OH
`
`HO
`
`OH
`
`Br
`
`Synthesis process includes the hydroxyl methylation, ester reduction, alcohol
`
`oxidation, witting reaction( or Homerreaction or Horner-Emmons-Wadsworth
`
`reaction) and demethylation. The synthesis routes are well established and available in
`
`the art.
`
`Additional stilbene derivatives may be obtained by standard esterification
`
`through the reaction of hydroxylated stilbene derivative and an acid orits derivative
`
`such as the correspondingsalt, chloride and anhydride. This reaction is well known in
`
`the art. For example, an alcohol was added to a mixture of anhydride and pyridine at
`
`low temperature, and the mixture wasleft at room temperature for sufficient time to
`
`complete the reaction. After the reaction, general work up process that is knownto the
`
`art gave the corresponding derivatives.
`
`20
`
`25
`
`The inventive compounds of formula I] may be produced with an appropriate
`
`oxidant, such as m-chloroperbenzoic acid, and hydrolysis of the corresponding
`
`compoundsof formula I. For example, 3,5-Dihydroxy-4-isopropyl-trans-stilbene
`
`epoxide can be synthesized from 3,5-dihydroxy-4-isopropyl-trans-stilbene compound
`
`(9) in Scheme 1 on next page by reacting with m-chloroperbenzoic acid (1.2 eg.) in
`
`CH2Chat 0°C. After the reaction, the general work up process is knownto the art. The
`
`compoundsofthe present invention mayalternatively be prepared from different routes
`
`reported in the literature and are incoporated herein as references. (Eicher, T.;
`
`Tiefensee, K.; Donig, R. and Pick, R. Synthesis 1991, 98-102; Krow, G. R.; Miles, W.
`
`H.; Smiley, P. M.; Lester, W. S. and Kim, Y. J. J. Org. Chem. 1992, 537, 4040-4043;
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`Seguineru, P. and Villieras 7 2trahedron Letters 1988, 29, 477-480; Thakkar, K.;
`
`Geahlen, R. L. and Cushman. M. J. Med. Chem. 1993, 36, 2950-2955; Bezou, P.;
`
`Hilberer, A. and Hadziioannou, G. Synthesis 1996, 449-451). These methodsare either
`
`using complex catalyst (Krow, G. R.; Miles, W. H.; Smiley, P. M.; Lester, W. S. and
`
`Kim, Y. J.J. Org. Chem. 1992, 57, 4040-4043) or involving manysteps (Eicher,T.;
`
`Tiefensee, K.; Donig, R. and Pick, R. Synthesis 1991, 98-102).
`
`(a)
`
`Schemes of Synthesis
`
`Compoundsofthe present inventions were synthesized from commercially
`
`available 3,4-dihydroxybenzoic acid (1) and 4-bromo-3,4-dihydroxybenzoic acid (10)
`
`following the route, outlined in Scheme1 or 2:
`
`COOMe
`
`COOMe
`
`CH,OH
`
`BBA,
`b-fioho ‘ Or
`
`_ wa
`
`20
`
`25
`
`(CH3)2SO04, CH;COCH;, K,CQ;, reflux, 24 h; ii. BrCH(CH;)2,
`Scheme1. i.
`AICh, CS), reflux 7d; iii. LiAIH4, Et,O, 0°C, 2h, ; iv. Py'CI'CrOs, 1.5 h; v.
`PO(OEt)3, (But),N*T, 110 -130 °C, 10 h; vi. NaH, epd 5, THF,60 °C, 3 h; vii.
`BBr3, CH2Ch, —78 °C then 0°C, 24 h.
`
`then 0°C, 24 h. R—- Primary alkyl groups, (-CH2CHs3, —(CH2):3CH3
`
`i. (CH3)2SQ0,, CH;COCH3, K2COs, reflux, 24 h; ii. LIALH4, Et,0,
`Scheme 2.
`0°C, 2h, ; iii. Py"CI-CrO3, 1.5 h; iv. NaH, 7, THF, 60 °C, 3 h; v. a. t-butyl
`Li, THF, —78 °C then to rt,2h, b. RBrin THF, vi. BBr3, CH)Ch, —78 °C
`
`SUBSTITUTE SHEET (RULE 26)
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`Procedure A (Methylation). Alcohol or acid (1 g, leq) was added to a well
`
`stirred acetone solution (100 ml) containing dimethyl sulfate (2 eq. for each hydroxyl or
`
`carboxylic group) and K2CQ;(5 eq. for each hydroxy] or carboxylic group). This
`
`solution was refluxed for 12 hours. After filtration, solvent acetone was evaporated
`
`under reduced pressure, the residue was dissolved in EtOAc (50 mL). The EtOAc
`
`solution was washed with water (50 mL x 2), saturated aqueous NaCl (50 mL), dried
`
`over Na?SOu, evaporated under reduced pressure to offer syrup which was purified by
`
`silica column chromatography (Hexanes/EtOAc = 4:1). When PO(OETt)3 is substituted
`
`with triphenylphosphate,cis-derivatives are synthesized.
`
`Procedure B (Reduction of methyl ester to alcohol). LiAIH,4 (1.5 eq.) was added
`
`slowly with stir to the methyl ester (1 g, 1 eq) in anhydrous diethyl cther (100 mL)at 0
`
`°C. After 30 min, water (5 mL) was added slowly to the mixture to quench excess
`
`LiAlHg, and the mixture wasacidified with 10 % HCI (aq). The organic layer was
`
`washed with water (50 mL x 2), saturated NaHCO; (50 mL), dried over Na2SOuand the
`
`solvent was evaporated under reduced pressure to offer syrup which wascrystallized
`
`from EtOH/hexanes.
`
`Procedure C (Oxidation of primary alcohol to aldehyde). Alcohol (1 g, 1 eq) in
`
`dichloromethane (10 mL) was addedto a well stirred anhydrous dichloromethane (50
`
`mL) containing suspended pyridinium chlorochromate (1.5 eq. to each hydroxyl
`
`group). After 90 min, diethyl ether (100 mL) was added, the supernatant was decanted
`
`and the black gummysyrup was washed with dry ether (20 mL x 3) and became a black
`
`solid. The combined organic solution was passed througha short pad of Florisol and
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`the solvent was removedbyrotary evaporation offering syrup that was then crystallized
`
`from EtOH/hexane.
`
`Procedure D (Wittig Reaction). NaH (2 cq.) was addedto a well stirred diethyl
`
`benzylphosphonate ester (7) (1.5 eq.) in dry THF (25 mL) for 60 min at 0 °C.
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`Aldehyde (1 g, 1 eq) in THF (2 mL) wasaddedslowly to the mixture, and the reaction
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`mixture was allowed to react for 3 hours at 50 °C. After cooling down to room
`
`temperature, the mixture was poured onto ice, followed by addition of 2M HCl (5 mL).
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`25
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`The mixture was extracted with EtOAc (20 mL x 3), the combined organic layer was
`
`then washed with water (25 mL x 2), saturated NaCl (25 mL) and dried over anhydrous
`
`magnesium sulfate. After filtration, the solvent was evaporated under reduced pressure,
`
`the resulting syrup was purified by silica column chromatographyy (Pet ether/ether =
`
`8:1).
`
`Procedure E (Demethylation): BBr3 (4 mL, 1 M in CH3Cly) in 10 mL of dry
`
`CHCl, was added dropwise into methylated stilbene (1 g, 1 eq) in dry CHiCh(5 mL)
`
`at — 78 °C, andleft at room temperature overnight. The mixture was then poured on the
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`ice and the organic layer was separated and aqueous layer was extracted with CH2Ch
`
`(20 mL x 2). The combined organic layer was washed with saturated NaCl, dried over
`
`anhydrous sodium sulfate and evaporated under reduced pressure to dryness.
`
`Procedure F (Acetylation). Alcohol (1 g,
`
`1 eq) was addedto an ice-cold
`
`mixture of acetic anhydride/pyridine = 1:1 (v/v) (5 mL), and the mixture was left at
`
`room temperature overnight. After the reaction, EtOAc (25 mL) was addedto the
`
`mixture, the EtOAc was washed with ice-cold water (25 mL), ice-cold 10 % HCI (25
`
`mL x 2), water (25 mL x 2), saturated aqueous NaHCO;(25 mL), and dried over
`
`anhydrous Na3SO4. After removal of the solvent the product was purified bysilica
`
`column chromatographyy (Hexanes/EtOAc = 8:1).
`
`(c}
`
`Use of Synthesis Scheme 1| to make 3,5-Dihydroxy-4-isopropyl-trans-stilbene
`
`andits derivatives.
`
`Gi) Methyl 3,5-dimethoxy-benzoate (compound 2 in Scheme 1) (Procedure A):
`
`the crude product (syrup) was crystallized from EtOH / Hexanesto give pure
`
`compound 2 (Scheme 1) (~ 90 %). Mp: 110-113 °C. NMR (100 MHz, CDC};) 6 4.20
`
`(s, 6 H, OCHS), 4.60 (s, 3 H, COOCHS), 6.30 (t, 1 H, J42 = 2.2 Hz, H-4), 7.20 (d, 2H,
`
`H-2,6).
`
`(11) Methyl 3,5-dimethoxy-4-isopropyl-benzoate (compound 3 in Scheme1):
`
`anhydrous AICI]; (0.85 g) was added to dry CS; (100 mL) containing methyl 3,5-
`
`dimethoxy-benzoate (compound 2 in Scheme 1) (0.86 g) and 2-bromopropane(0.61
`
`mL, 1.1 eq.). This solution was heated to reflux for 7 days. The mixture wasfiltered,
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`20
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`25
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`washed with water (100 mL x 2), saturated NaHCO; (100 mL) and saturated NaCl (100
`
`mL), dried over Na2SOg,. Afterfiltration and removal of the solvent, the crude product
`
`waspurified by column chromatographyy (EtOAc/Hexanes = 4:1) to give methyl 3,5-
`
`dimethoxy-4-isopropyl-benzoate (compound 3) (0.69 g, 66 %) which wascrystallized
`
`from EtOH/Hexanes. NMR (100 MHz, CDCIs) 8 1.61 (d, 6 H, Jy.2 = 7.1 Hz, H-2’),
`
`3.66 (hept, | H, H—-1’), 3.88 (s, 6 H, OCH3), 3.94 (s, 3 H, COOCH3), 7.25 (s, 2 H, H-
`
`2,6).
`
`(111) 3,5-Dimethoxy-4-isopropyl-benzyl alcohol (compound4 in Scheme 1)
`
`(Procedure B): The crude product (syrup) wascrystallized from EtOAc/Hexanesto
`
`give compound 4 (85 % yield). NMR (100 MHz, CDC1;) 6 1.31 (d, 6 H, Jy.2.= 7.1 Hz,
`
`H-2'), 3.61 (hept, 1 H, H-1’), 3.84 (s, 6 H, OCH3), 4.68 (s, 1 H, H-OH), 6.60 (s, 2 H,
`
`H-2,6).
`
`(iv) 3,5-Dimethoxy-4-isopropyl-benzaldehyde (compound 5 in Scheme 1)
`
`(Procedure C): The resulting residue was crystallized from EtOH to give pure
`
`compound 5 (80 % yield). NMR (100 MHz, CDCI) 6 1.31 (d, 6 H, Jy.2 = 7.1 Hz, H-
`
`2"), 3.61 (hept, 1 H, H-1’), 3.84 (s, 6 H, OCHS), 4.68 (s, | H, H-OH), 6.60 (s, 2 H, H-
`
`2,6).
`
`(v) Diethyl benzylphosphonate ester (compound 7 in Scheme1): Triethyl
`
`phosphite (3.2 mL, 1.5 eq.) was added to the benzyl bromide (2.11 g) containing a
`
`catalytic amount of tetrabutylammonium iodide, the mixture was heated to 110-130 °C
`
`overnight. Excess triethyl phosphite was removed by heating the solution for | hour at
`
`100 °C under vacuum (15 mm Hg)to yield compound 7 quantitatively. NMR (100
`
`MHz, CDC1s) 6 1.21 (t, 6 H, Joy = 7.1 Hz, H-2’), 3.02 (s, 1 H, H-a), 3.24 (8,
`
`1 H, H-
`
`B), 3.98 (quint, 2 H, H-1'), 7.27 (s, 5 H, H-ArH).
`
`(vi} 3,5-Dimethoxy-4-isopropyl-trans-stilbene (compound 8 in Scheme 1)
`
`(Procedure D): The product was purified by column chromatography (Et2O/Hexanes =
`
`1:8) and crystallized from cther/hexanes to give pure compound 8 (70 % yield). Mp:
`
`64-66 °C. NMR (400 MHz, CDCI) 6 1.28 (d, 6 H, Imecu= 7.0 Hz, CHa), 3.58 (hept, 1
`
`20
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`H, -CH_), 3.85 (s, 6 H, H-OCHs3), 6.69 (s, 2 H, H-2, 6), 7.05 (s, 2 H, =CH), 7.25 (m, 1
`
`H, H-4’), 7.35 (m, 2 H, H-3’,5’), 7.25 (m, H-2’,6’).
`
`(vii) 3,5-Dihydroxy-4-isopropyl-irans-stilbene (compound 9 in Scheme1)
`
`(Procedure E): The product was purified by column chromatography (EtOAc/Hexanes)
`
`and give desired compound 9 (95 % yield). Mp: 140-142 °C. NMR (400 MHz, CDCl;)
`
`5 1.38 (d, 6 H, Ja.g= 7.3 Hz, CH), 3.46 (hept, 1 H, -CH-), 4.80 (s, 2 H, H-OH), 6.50
`
`(s, 2 H, H—2, 6), 6.92 (d, 1 H, Jap = 16.2 Hz, H- =CHa), 6.97 (d, 1 H, H-=CHp), 7.25
`
`(m, 1 H, H-4'), 7.34 (m, 2 H, H-3’,5’), 7.52 (m, 2 H, H-2’,6’).
`
`(d) Use of Scheme2 to form additional 4-substituted 3,5-dihydroxy-trans-stilbenes and
`
`their derivatives.
`
`(vill) Methyl 3,5-dimethoxy-4-bromo-benzoate (compound 11 in Scheme2)
`
`(Procedure A). The crude product (95 %) was crystallized from EtOH/ Hexanes. Mp:
`
`119-124 °C. NMR(100 MHz, CDCl) 6 3.96 (s, 3 H, COOCHS), 3.99 (s, 6 H, OCHS),
`
`7.28 (s, 2 H, H-2, 6).
`
`(ix) 3,5-Dimethoxy-4-bromo-benzyl alcohol (compound 12 in Scheme 2)
`
`(Procedure B). The crude product (85 % ofyield) was crystallized from EtOQH/
`
`Hexanes.Mp: 84-95 °C. NMR (100 MHz, CDCl) 6 1.95 (s, 1 H, OH), 3.93 (s, 6 H,
`
`OCH3), 4.69 (s, 1 H, CHz), 6.61 (s, 2H, H-2,6).
`
`(x) 3,5-Dimethoxy-4-bromo-benzaldehyde (compound 13 in Scheme 2)
`
`(Procedure C). The crude product (75 %) was crystallized from EtOH/Hexanes. Mp:
`
`110-113 °C. NMR (100 MHz, CDC1s) 6 4.02 (s, 6 H, OCH3). 7.11 (s, 2H, H-2,6), 9.97
`
`(s, 1 H, CHO).
`
`(xi) 3,5-Dimethoxy-4-bromo-trans-stilbene (compound 14 in Scheme 2)
`
`(Procedure D). The crude product was purified by column chromatography (Pet
`
`ether/Ether = 8:1) in 70 % and crystallized from ether/hexanes. Mp: 149-152 °C.
`
`NMR(400 MHz, CDC]s) 8 3.96 (s, 6 H, 2 x OCHs), 6.72 (s, 2 H, H—2,6), 7.06 (d, 1 H,
`
`10
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`20
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`25
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`30
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`SUBSTITUTE SHEET (RUL