Leading bulk Producers of "withaferin A" "Now avilable in all quantities up to 1000 grams level"
     PHARMACOLOGICAL ACTIVITIES OF WITHAFERIN A

pharmacological activities of withaferin a

 

 

Withaferin A has shown significant anti-cancer activity in animal studies

Withaferin A has been extensively studied for

  • Anti- inflammatory
  • Anti- tumor
  • Anti-angiogenesis
  • Radio sensitizing activity
  • Chemo preventive and
  • Immunosuppressive activities
  1. ANTI-INFLAMMATORY ACTIVITY:
  2. The anti-inflammatory effect of withaferin A on a experimental mice model for gouty arthritis monosodium with urate crystal induced inflammation was studied. A cellular model of cystic fibrosis inflammation was established by Maitra et al to assess the anti-inflammatory activity of withaferin A 1,2

  3. ANTI-TUMOR AND RADIOSENSITIZING ACTIVITY:
  4. Shohat et al first reported the anti-tumor activity of withaferin A in 1967 3. They also studied the effect of withaferin A on Ehrlich ascites tumor cells 4, 5. Sharada et al confirmed the anti-tumor effect of withaferin A against mouse Ehrlich ascites carcinoma cells. In addition, radiosensitizing effect of withaferin A was first demonstrated and an optimum dose of 30 mg/kg withaferin A in combination with 7.5 Gy gamma radiation for Ehrlich ascites implanted mice was demonstrated by Devi et al 6,7. The anti-tumor and radiosensitizing activities of withaferin A were tested for different cancer cell lines. Devi et al evaluated the radiosensitizing effect of withaferin A on B16F1 mouse melanoma cells and mouse fibrosarcoma grown in C57B1 and swiss albino mice8,9. Kalthur et al confirmed that withaferin A, hyperthermia and irradiation acted synergistically against B16F1 melanoma and withaferin A served as a better radio sensitizer than hyperthermia 10. 
    The anti-tumor effect of withaferin A against human prostate cancer cell lines was tested. Withaferin A exhibited androgen receptor (AR) dependent cell killing against prostate cancer cell lines 11. It was confirmed in PC-3 xenografts in nude mice. Withaferin A was also demonstrated to inhibit HSP 90 by directly binding to the C-terminus, which was different from classical HSP 90 inhibitors and thus proposing a new mechanism for developing HSP 90 inhibitors. Combined withaferin A with myricetin enhanced anti-cancer efficacy in pancreatic cancer cells. Withaferin A 6 mg/kg inhibited tumor growth in pancreatic panc-1 xenografts 11A.
    Stan et al demonstrated that withaferin A inhibited proliferation of breast cancer cells with IC 50 s of 1.5 µm for MCF-7 cancer cells and 2.0µm for MDA-MD-231 cancer cells12. 
    Withaferin A exhibits anti-proliferative activity against human promyelocytic leukemia cells HL-60, U937, lymphoid origin human T-(MOLT-4, Jurkat), B-(REH) cells and myeloid origin K-562 leukemic cells, but not the normal lymphocytes peripheral blood mononuclear cells(PMC)13,14,15.
    Withaferin A inhibited cell survival in three human colon cancer cell lines SW-480, SW-620 and HCT-116 without affecting normal colon epithelial FHC cells 16. 
    The anti-proliferative activity of withaferin A was demonstrated against human head and neck squamous cell carcinoma UM-SCC-2, MDA 1986, JMAR & JHVO 11 17.

  5. ANTI-ANGIOGENESIS ACTIVITY:
  6. Anti-angiogenesis activity of withaferin A was first demonstrated by Mohan et al 18. Withaferin A was shown to inhibit human umbilical vein endothelial cell (HUVEC) sprouting in three –dimensional collagen-1 matrix, inhibit HUVEC cell proliferation with IC50 of 1.2 nM and exert potent anti-angiogenic activity in FGF-2 matrigel plug angiogenesis mice model at doses as low as 7µg/kg/day which were 500 fold lower than the reported doses to exert anti-tumor activity in vivo. Withaferin A significantly inhibited neovascularization in injury-induced carneal neovascularization mouse model by about 70% 19.

  7. CHEMOPREVENTIVE ACTIVITY:
  8. Chemopreventive role of withaferin A was demonstrated. It was shown that 7, 12-dimethyl benz[a] anthracene (DMBA) induce oral carcinogenesis in syrian golden hamsters, Where as oral administration of 20 mg/kg withaferin A could completely prevent the tumor induction by DMBA 20, 21.

References:

  • Sabina et al, Inhibition of monosodium urate crystal-induced inflammation by withaferin A. J. Pharm Pharm Sci 2008; 11: 46-55
  • Maitra et al, Inhibition of NF kappa B by the natural product withaferin A in cellular models of cystic fibrosis inflammation. J inflamm (London)  2009; 6 :15
  • Shohat et al, Antitumor activity of withaferin A (NSC 101088). Cancer chemotherapy reports  1967; 51; 271-276
  • Shohat et al, Effect of withaferin A on Ehrlich ascites tumor cells-cytological observations. Int J Cancer  1970; 5: 244-252
  • Shohat et al, Effect of withaferin A on Ehrlich ascites tumor cells. II. Target tumor cell destruction in vivo by immune activation.  Int J Cancer 1971; 8 ; 487-496
  • Devi et al, In vivo growth inhibitory and radiosensitizing effects of withaferin A on mouse Ehrlich ascites carcinoma. Cancer Lett 1995; 95: 189-193
  • Sharada et al, Antitumor and radiosensitizing effects of withaferin A on mouse Ehrlich ascites carcinoma in vivo. Acta Oncol  1996; 35 : 95-100
  • Devi et al, Radiosensitization of a mouse melanoma by withaferin A in vivo studies. Indian J Exp Biol 2000; 38: 432-437
  • Umadevi et al, Radiosensitizing effect of withaferin A combined with hyperthermia on mouse fibrosarcoma and melanoma. J Radiat Res (Tokyo) 2003; 44 : 1-6 
  • Kalthur et al, Enhancement of the response of B16F1 melanoma to fractionated radiotherapy and prolongation of survival by withaferin A and/or hyperthermia. Integr Cancer Ther  2010; 9: 370-377
  • Srinivasan et al, Par-4-dependent apoptosis by the dietary compound withaferin A in prostate cancer cells. Cancer Res  2007; 67; 246-253, (11A). Yanke yu , Ph.D Thesis, 2011. Pharmaceutical sciences, The University of Michigon, USA.
  • Stan et al, Withaferin A causes FOXO3a and Bimdependent apoptosis and inhibits growth of human breast cancer cells in vivo. Cancer Res 2008; 68: 7661-7669
  •  Malik et al, Reactive oxygen species generation and mitochondrial dysfunction In the apoptic cell death of human myeloid leukemia HL-60 cells by a dietary compound withaferin A with concomitant protection by N-acetyl cysteine. Apoptosis 2007; 12: 2115-2133
  • Oh et al, Induction of apoptosis by withaferin A in human leukemia U 937 cells through down-regulation of Akt phosphorylation. Apoptosis 2008; 13; 1494-1504
  • Mandal et al, Withaferin A induces apoptosis by activating P38 mitogen-activated protein kinase signaling cascade in leukemic cells of lymphoid and myeloid origin through mitochondrial death cascade. Apoptosis 2008; 13: 1450-1464
  • Koduru et al, Notch-1 inhibition by withaferin A ; a therapeutic target against colon carcinogenesis. Mol Cancer Ther 2010; 9: 202-210
  • Samadi et al, Withaferin A a cytotoxic steroid from vassobia breviflora, induces apoptosis in human head and neck squamous cell carcinoma. J. Nat Prod 2010; 73: 1476-1481
  • Mohan et al, Withaferin A is a potent inhibitor of angiogenesis. Angiogenesis 2004; 7: 115-122
  • Barganag Mohan et al, The tumor inhibitor and anti-angiogenic agent withaferin A targets the intermediate filament protein vimentin.  Chem Biol 2007; 14: 623-634
  • Manoharan et al, Circadian time-dependent chemopreventive potential of withaferin A in 7, 12-dimethylbenz[a] anthracene induced oral carcinogenesis. Pharmacol Rep 2009; 61: 719-726
  • Manoharan et al, Protective effect of withaferin A on tumor formation in 7,12-dimethylbenz[a] anthracene induced oral carcinogenesis in hamsters. Ind j Exp Biol  2009; 47: 16-23

STRUCTURE ACTIVITY RELATIONSHIP OF WITHAFERIN A AND ITS PHARMACOLOGICAL ACTIVITY:
The key structural components in withaferin A which contribute to its biological activities were elucidated.
Withaferin A is highly reactive towards proteins as the ketone containing unsaturated A ring (Double bond at C2-C3 Position), the epoxide ring within B ring and unsaturated lactone ring were all demonstrated to be involved in Michael addition, thioalkylation reactions. The double bond in ring A is crucial for the cytotoxicity of withaferin A. The double bond dissociated derivatives of withaferin A showed little cytotoxicity. Where as the C27 hydroxyl group and the α,β-unsaturated δ-lactone ring (Double bond at C24-C25) are not required for the cytotoxicity of withaferin A. 5β,6β-epoxide ring is crucial for its cytotoxicity .  The conjugated ketone carbon in the ring A is required for the proteasome inhibition. The C2-C3 unsaturated position of the A ring and C5-C6-epoxide group contributes to the binding of withaferin A to vimentin. Whereas withaferin A derivatives 3β-methoxydihydrowithaferin A (or) thiophenoxydihydrowithaferin A (Without C2-C3 double bond) failed to inhibit vimentin 1-5.

REFERENCES:

    • Fuska et al, Novel cytotoxic and anti-tumor agents. Withaferin A relation of its structure to the in vitro cytotoxic effects on P-388 cells.  Neoplasma  1984; 31: 31-36
    • Yokota et al, development of withaferin A analogues as probes of angiogenesis.   Bioorg Med Chem Lett  2006; 16: 2603-2607
    • Damu et al, Isolation, structures and structure cytotoxic activity relationships of withanolides and physalins from physalis angulata. J.Nat Prod  2007; 70: 1146-1152
    • Yang et al, The tumor proteasome is a primary target for the natural anti-cancer compound withaferin A isolated from “Indian winter cherry”.  Mol Pharmacol  2007; 71: 426-437
    • Bargagna-Mohan et al, The tumor inhibitor and anti- angiogenic agent withaferin A targets the intermediate filament protein vimentin.  Chem Biol  2007; 14: 623-634

STRUCTURAL MODIFIED DERIVATIVES OF WITHAFERIN A:
Yokota et al developed a biotinylated affinity analogue of withaferin A for use as a probe to study angiogenesis1.
A library of 2, 3-dihydro-3β-substituted withaferin A derivatives were prepared by regio/stereoselective Michael addition to ring A.  The analogues were tested for in vitro cytotoxicity against various human cancer cell lines. 3- Azido analogue exhibited 35 fold increase in cytotoxicity against all cell lines compare to parent molecule2.
A series of withaferin A analogues were prepared and tested for its activity in proliferative diseases, neurodegenerative diseases, autoimmune and inflammatory diseases3.

REFERENCES:

    • Yokota et al, development of withaferin A analogues as probes of angiogenesis.   Bioorg Med Chem Lett  2006; 16: 2603-2607
    • Yousuf et al, Ring A structural modified derivatives of withaferin A and the evaluation of their cytotoxic potential.  Steroids  2011; 76:  1213-1222
    • Gunatilake et al, Withaferin A analogues and uses thereof. US patent 2011/023055A1

    OTHER IMPORTANT REFERENCES:

    1. Kaileh et al, Withaferin A strongly elicits lkappaB kinase beta hyperphosphorylation concomitant with potent inhibition of its kinase activity. J Biol Chem 2007; 282: 4253-4264
    2. Umadevi et al, Enhancement of radiation induced cell death in chicken B lymphocytes by withaferin A. Indian J Exp Biol 2008; 46: 437-442
    3. Shohat et al, Immunosuppressive activity of two plant steroidal lactones withaferin A and withanolide E. Biomedicine 1978; 28: 18-24
    4. Devi et al, Withaferin A a new radiosensitizer from the Indian medicinal plant withania somnifera. Int j Radiat Biol 1996; 69; 193-197
    5. Sen et al, Apoptosis is induced in leishmanial cells by a novel protein kinase inhibitor withaferin A and is facilitated by apoptotic topoisomerage I-DNA complex. Cell Death Differ 2007; 14: 358-367
    6. Falsey et al, Actin microfilament aggregation induced by withaferin A is mediated by annexin II. Nat Chem Biol 2006; 2: 33-38
    7. Lee et al, Withaferin A sensitizes Trail-induced apoptosis through reactive oxygen species-mediated up-regulation of death receptor 5 and down regulation of c-FLIP. Free Radic Biol Med 2009; 46: 1639-1649
    8. Lee et al, Withaferin A inhibits activation of signal transducer and activator of transcription 3 in human breast cancer cells. Carcinogenesis 2010; 31: 1991-1998
    9. Liang et al, Inhibition of transcription factor NF-KappaB signaling proteins IKK beta and P65 through specific cysteine residues by epoxyquinone A monomer correlation with its anti-cancer cell growth activity. Biochem Pharmacol 2006; 71: 634-645
    10. Oh et al, Withaferin A inhibits i NOS expression and nitric oxide production by Akt activation and down-regulating LPS –induced activity of NF-KappaB in RAW 264.7 cells. Eur J Pharmacol 2008; 599: 11-17
    1. Kuroyanagi et al, Cell differentiation inducing steroids from withania somnifera L. Chem Pharm Bull   1999; 47: 1646-1649



    DEVELOPED A PATENTED ( Indian Pat. Appl. (2012), IN 2011CH00055 A 20121019 ) PROCESS TO ISOLATE BULK QUANTITIES OF WITHAFERIN A. WE CAN MEET ANY QUANTITIES OF INDUSTRIAL REQUIREMENTS WITH PURITIES >98% BY HPLC.FOR FURTHER DETAILS PLEASE FEEL FREE TO CONTACT withaferina@gmail.com / xenonbiosciences@gmail.com (OR) +91-9849601776
.




.

One of the withaferin A derivative has been emerged as a preclinical lead for Amyotrophic Lateral Sclerosis (ALS).

Method of achieving normoglycemia in diabetics by administration of Withaferin A, US 8637494 B2

Molecule of the Week

Withaferin A Shows Early Promise to Treat Pancreatitis