Human Clinical Studies

Wei S, Qiu L; Chen ZH; Xu MC. Effect observation of Snake venom point injection in the treatment of scapulohumeral periarthritis. Chinese Journal of Information on Traditional Chinese Medicine, 2007. 14: 68，96.

Xu JM, Song ST, Feng FY, Huang FL, Yang Y, Xie GR, Xu LG, Paradiso A. Compound Keluoqu for treatment of chronic moderate to severe cancer pain: a multicenter randomized trial. ASCO Annual Meeting (2002) Abstract No. 1762

Xu JM, Song ST, Yu1 CZ, Yang Y, Cui M, Wang WU, Xiong YL, Paradiso A Randomized, Double-blind, Placebo-controlled, Parallel Multicenter Trial of Cobrotoxin-containing Compound Analgesic in the Treatment of Postoperative Pain. Europain (2002) 2, Issue 2, Abstract

Wu YQ, Yu W. Clinical observation of capsule in the treatment of infertility group in 122 cases of the digestive system cancer. Journal of Snake, 2001. 13: 26-28.

Wei S, ZW Huang. Snake venom injection point injected in the treatment of altauna by rheumatoid arthritis in 32 cases. Journal of Shandong University of Traditional Chinese Medicine, 2000. 24: 276-277.

Wang X, Wang F, and Hu ZD, Effect of new Cobratoxin on Postoperative analgesia, J. Snake 1999; 11 (1): 19-20

Wu MB, Wu GL. The treatment of rheumatoid arthritis in 126 cases of venom compound capsule. Journal of Snake, 1999, 11: 30.

Zhu T, Liu G, Ji T, Miao Z and Chen S. The Analgesic effect of injectional cobra neurotoxin purified judged by the National Standard in Clinic. J. Snake 1999; 11 (4): 43-47

Gao ZN. Analysis on the therapeutic effect of Cobrotoxin with Chinese traditional medicine for sciatica in 182 cases. Journal of Snake, 1998. 10: 21.

Zhu CZ,Li H, Yang QP, et al. The clinical treatment of diabetic kidney capsule of Cobra venom. Medicine and Pharmacy of Yunnan, 1997. 18: 460-461

Wang SN, Wang H, Peng SF, Zhang HL. The current situation in research and clinical application of the cobra venom. Journal of Snake, 1996, 08: 23-27.

Cao YCB and Zhao GCY, KTL Clinical Anlgesic effect J. Snake 1995; 7, No. 1: 51-53

Cao YS, Cheng BQ, Zhao GH, Chen YM. Analgesic effect of Ke Ning Compound pain in clinical application. Journal of Snake, 1995, 7: 51~53.

Zhou LH, Liu MJ, Liu ZL et al. The effect on hemorheology of snake venom ingredient M. Journal of Chinese Microcirculation, 1993: 79-80.

Xiong Y, Wang W, Pu X, Song, Liu L, and Mao J, Using snake venom to substitute for addictive drugs, Toxicon 1992; 30:567-568

Li DH, Zhou YL. Clinical Observation on Treatment of the Cobrotoxin in 90 cases with heroin dependence. Medicine and Pharmacy of Yunnan. 1991, 12: 322-324.

Song YT. Clinical observation of snake venom on lung cancer. Clinical. 1991. 6: 311-312.

Wang S, He Y, Peng S. Preparation of venom and its analgesic activity. Zhongcaoyo 1986 17(1): 7-9

Pu DX. Observation on the therapeutic effect of Cobrotoxin for neurovascular headache, sciatica and trigeminal neuralgia in 96 cases. Medicine and Pharmacy of Yunnan, 1980: p. 27-31

Zeng CM. Observation on the therapeutic effect of the novel drug "Cobrotoxin" for chronicity backleg pain in 80 cases. Medicine and Pharmacy of Yunnan, 1980: p. 26-28.

Wenshan state dermatosis institute, Yunnan Province, A clinical observation report on the novel drug "Cobrotoxin" for reactivity neuralgia of leprosy. Journal of Dermatology and Venereology, 1978: 03-05.

Singh I, Srivastava MC. Anti-asthmatic effects of immunization with cobra venom. Br J Dis Chest. 1968 Apr;62(2):107-9.

Williams EY. Treatment of trigeminal neuralgia with cobra venom. J Natl Med Assoc. 1960 Sep;52:327-8.

Meiselas LE, Schlecker AA. The effect of Nyloxin on the pain of arthritis. N Y State J Med. 1957 Jun 15;57(12):2067-8.

Teichman J, Cobratoxin and its use in medicine, Wien Med Wochenschr. 1956 Apr 21;106(16):364-7.

Bryson KD. The treatment of chronic arthritis with a combination of cobra venom, formic acid, and silicic acid. Am Surg. 1954 Jul;20(7):751-5.

Jackman AI. Cobra venom therapy in the neuroses; a preliminary report. Dis Nerv Syst. 1954 Apr;15(4):99-102.

Lumpkin WR, Firor WM, Evaluation of the Bryson treatment of arthritis. Am. Surgeon (1954); 20:756-759

Oaks LW, Quinn JH. Cobra venom in ocular therapy. Trans Pac Coast Otoophthalmol Soc Annu Meet. 1954 35:71-82.

Taren JA. A clinical evaluation of cobra venom extracts for control of pain. Med Bull (Ann Arbor). 1953 Aug;19(8):206-11.

Grasset E. [The cobra neurotoxin; pharmacology and clinical applications in the treatment of pain.], Med Hyg (Geneve). 1952 Feb 15;10(212):55-8.

Hills RG, Firor WM. The use of more potent cobra venom for intractable pain. Am Surg. 1952 Sep;18(9):875-9.

Seliger H. [New results with cobratoxin in cancer therapy.] Med Welt. 1951 May 12;20(19):638.

Tan MG and Ines-Tan Ar, Treatment of Herpes Zoster with cobra venom and sulfanilamide: report of 5 cases, J Philipp Med Assoc. 1947 Dec;23(12):593-9.

Black WT. Cobra venom for the relief of pain. Sout. M. J. 1940; 33: 432

Hayman M, Macht DI. Clinical and biochemical studies in cobra venom therapy. Med. Rec. 1940; 152: 67

Macht DI. New developments in pharmacology and therapeutics of cobra venom. Tr. Am. Therap. Soc. 1940; 40

Rutherford RN, The Use of Cobra Venom in the Relief of Intractable Pain, NEJM 1939: 408-413

Gayle, RF, and Williams, JN, Symptomatic treatment of Parkinsonism symptoms with cobra venom, South. M. J. 1938; 31: 188-192,

Macht DI, Therapeutic experiences with Cobra venom. Ann. Int. Med. 1938; 11: 1824•1833.

Kirchen M, Uber das Kobratoxin und seine therapeutische Verwendung, Wien. Klin. Wehnschr 1936; 49: 648-651.

Macht DI. Experimental and Clinical Study of Cobra Venom as an Analgesic. Proc Natl Acad Sci U S A. 1936 Jan;22(1):61-71.

Van Esveld LW. Preparation of “Cobratoxin” for clinical purposes, especially for the treatment of cancer pains, Biochemische Zeitschrift 1936; 283: 343-57

Laignel-Lavastine, Korossios NT, Cobra Venom Therapy. J. Med. Paris, 1933; Vol. LIII: 652.

Taguet C, La cure des algies et des tumeurs malignes. Bull. et mem Soc. de Med. de Paris 1933; 131: 310.

Monaelesser A, and Taguet C, Traitment des algies et des tumeurs par le “Venin du Cobra” Bull. Act. De Med Paris 1933; 109: 371-377

Pre-clinical Research

Catassi A, Paleari L, Servent D, Sessa F, Dominioni L, Ognio E, Cilli M, Vacca P, Mingari M, Gaudino G, Bertino P, Paolucci M, Calcaterra A, Cesario A, Granone P, Costa R, Ciarlo M, Alama A, Russo P. Targeting alpha7-nicotinic receptor for the treatment of pleural mesothelioma. Eur J Cancer. 2008 Oct;44(15):2296-311.

Grozio A, Paleari L, Catassi A, Servent D, Cilli M, Piccardi F, Paganuzzi M, Cesario A, Granone P, Mourier G, Russo P. Natural agents targeting the alpha7-nicotinic-receptor in NSCLC: a promising prospective in anti-cancer drug development. Int J Cancer. 2008 Apr 15;122(8):1911-5.

Chen R, Robinson SE. Effect of cholinergic manipulations on the analgesic response to cobrotoxin in mice. Life Sci 1990; 47: 1949-1954.

Chen ZX, Zhang HL, Gu ZL, Chen BW, Han R, Reid PF, Raymond LN, and Qin ZH. A long-form α-neurotoxin from cobra venom produces potent opioid independent analgesia. Acta Pharmacologica Sinica 2006; 27: 402-8.

Cheng BC, Zhou XP, Zhu Q, Gong S, Qin ZH, Reid PF, Raymond LN, Yin QZ, Jiang XH. Cobratoxin inhibits pain-evoked discharge of neurons in thalamic parafascicular nucleus in rats: Involvement of cholinergic and serotonergic systems. Toxicon. 2009 Apr 16.

Chopra RN, Chowhan JS, Snake venoms in Pharmacology and Therapeutics Ind. Med. Gaz. 1935; 445-453

Lamb G, Hunter W. On the action of venoms of different species of poisonous snakes on the nervous system. Lancet 1904 Jan 2; 1: 20

Liu YL, Lin HM, Zou R, Wu JC, Han R, Raymond LN, Reid PF, Qin ZH. Suppression of complete Freund's adjuvant-induced adjuvant arthritis by Cobratoxin. Acta Pharmacol Sin. 2009

Macht DI. Comparison of cobra venom and morphine as analgesics. Proceeding 1936; 101

Macht DI, Effects of repeated injections of cobra venom on blood chemistry and morphology, Proc. Soc. Exp. Biol Med. 43; 450 (1940)

Pu XC, Wong PT, Gopalakrishnakone P. A novel analgesic toxin (hannalgesin) from the venom of king cobra (Ophiophagus hannah). Toxicon 1995; 33: 1425-1431.

Zhang, HL, Han R, Gu ZL, Chen ZX, Chen BW, Reid PF, Raymond LN, Qin ZH, A short-chain a-neurotoxin from Naja naja atra produces potent cholinergic-dependent analgesia. Neuroscience Bulletin March 2006; 22(2): 103-110

Davenport HW. Prevention and suppression by azathioprine of venom-induced protein-losing gastropathy in dogs. Proc Natl Acad Sci U S A. 1976 Mar;73(3):968-70.

Ismail M, Aly MH, Abd-Elsalam MA, Morad AM. A three-compartment open pharmacokinetic model can explain variable toxicities of cobra venoms and their alpha toxins. Toxicon. 1996 Sep;34(9):1011-26.

Lee CY, Tseng LF. Distribution of Bungarus Multicinctus Venom Following Envenomation. Toxicon, 1966; 3: 281-290

Tseng LF, Chiu TH, Lee CY. Absorption and Distribution of I-Labeled Cobra Venom and Its Purified Toxins. Tox. And Applied Pharmac., 1968; 12: 526-535

Pittman R., Oppenheim R.,W: “Cell death of motoneurons in the chick embryo spinal cord. IV. Evidence that a functional neuromuscular interaction is involved in the regulation of naturally occurring cell death and the stabilization of synapses.” J Comp Neurol. 1979 Sep 15;187(2):425-46

Levin ME, Jin JG, Ji RR, Tong J, Pomonis JD, Lavery DJ, Miller SW, Chiang LW. Complement activation in the peripheral nervous system following the spinal nerve ligation model of neuropathic pain. Pain. 2008 Jul;137(1):182-201.

Jiang WJ, Liang YX, Han LP, Qiu PX, Yuan J, Zhao SJ. Purification and characterization of a novel antinociceptive toxin from Cobra venom (Naja naja atra). Toxicon. 2008 Oct;52(5):638-46

Xiong Y, Wang W, Pu X, Song, Liu L, and Mao J, Using snake venom to substitute for addictive drugs, Toxicon 1992; 30:567-568

Mechanism of Action

Pharmacology

The principal active components in the venoms are neurotoxins. These neurotoxins primarily target the cholinergic system by blocking the activity of acetylcholine, and compete with nicotine for receptor binding proving they are nicotinic antagonists. Nicotinic receptors are widespread throughout the body and are present on a variety of cells including nerve, immune and muscle. By targeting nicotinic receptors cobra venoms and cobra neurotoxins have recently been shown to have anti-inflammatory, antitumor and analgesic activity. The literature suggests that these activities are most likely mediated through 3 of the 10 known nicotinic receptors; alpha1, alpha7 and alpha9.

Pharmacodynamics

Characteristically, the onset of pharmacodynamic activity of peripherally administered cobra venom and cobra toxins is realized only after several hours in contrast to aspirin and morphine but the activity is more prolonged. In animal models Nyloxin does not induce tolerance nor addiction.

Constant administrations orally or by injection of small dosage of cobra neurotoxins increase the Leu-enkephalin content in hypothalamus, striatum and midbrain and increase the Met-enkephalin content in hypothalmus and midbrain, especially thalamencephalon. Cobratoxin exhibited a dosedependent analgesic action during Phase 1 and Phase 2 cycles in the formalin model. In this model formalin increased the number of c-Fos-positive cells in the L4-5 spinal dorsal horn. Peripheral treatment with Cobratoxin inhibited formalin-induced increases in c-Fos-positive cells, atropine (5mg/Kg) antagonized the antinociceptive activity and canceled the inhibitory effect of Cobratoxin on c-Fos expression. Najanalgesin, a newly isolated peptide neurotoxin from cobras with homology to Cardiotoxins, displayed analgesic activity with similar pharmacodynamics to the nicotinic antagonists, with slow onset, prolonged activity and antagonism by atropine. NTXI, a small peptide, could strikingly increase the pain threshold of mice in the hot plate assay. Intraperitoneal injection of 0.2 mg/kg of NTXI could increase pain threshold from 100% to 184.35% in mice on the hot plate’s threshold. The onset of the analgesic effect was slow, starting 2h after treatment reaching its maximal effect after 4 h. In addition to neurotoxins, it should be noted that cobra venom factor (CVF) also modulates the immune system through the complement cascade and has been reported to be effective in neuropathic pain models, which confirmed the importance of the immune system in the neuropathic pain process.

Cobra venom was recently reported to ameliorate adjuvant-induced arthritis in rats not only reducing pain but also reducing tissue damage confirming it as a potential disease modifying anti-rheumatic drug. Native cobra venom therapy at two different doses showed a significant protection against adjuvant-induced arthritic changes in several symptoms such as paw weight, paw and ankle diameters. The restoration of urinary hydroxyproline by cobra venom treatment indicated that it inhibited collagen break down and thereby prevented the cartilage damage caused by disease induction. Cobra venom treatment also significantly restored urinary glucosamine level. In adjuvant induced arthritis the alteration of glycohydrolase activity in joints may increase excretory glucosamine level. Thus restoration in excretory glucosamine concentration by the administration of cobra venom may protect against cartilage degradation. In the present study it was found that the levels of IL-10 were decreased in arthritic animals and venom treatment significantly increased serum IL-10 level. Cobratoxin was recently confirmed to have anti-inflammatory effects in the rat formalin and adjuvant arthritis models. Cobratoxin also induced changes in the expression of Th1 inflammatory cytokines and up-regulating the expression of the Th2 cytokine IL-10 thereby establishing dual mechanistic pathways for analgesic and anti-inflammatory activity, similar to the effects of whole cobra venom. Cobratoxin also appeared to protect joints from inflammatory damage. It has been reported that cobra venom also displayed anti-inflammatory activity in several animal models such as Xylene induced ear edema in mice, rat air pouch model and carrageenan induced paw edema model.

Cobra venom contains small amounts of other peptides that block muscarinic receptors, calcium and voltage-activated potassium channels, inhibitors of the complement cascade in addition to nerve growth factors.