Methods of combating snake toxin effects

Abstract

A method of combating snake toxin effects comprising administering to an individual in need thereof a pharmaceutical composition comprising a class of flavonoid compounds. The flavonoid compound is selected from a group consisting of isorhamnetin, quercetin and a combination of the isorhamnetin and the quercetin.

Description

Technical Field

[0001] The present disclosure relates to a method of using a pharmaceutical composition containing a flavonoid compound to counteract the effects of snake venom. Background Art

[0002] Snake venom is a highly toxic saliva found in various snake species and contains a variety of proteins and enzymes that are responsible for its toxicity. It is generally divided into two types, both of which are highly lethal to the victim. One is hemotoxic venom, which is found in Protobothrops spp. (such as Protobothrops mucrosquamatus), and the other is neurotoxic venom, which comes from Naja spp. (such as Naja nivea and Naja atra). When dealing with an emergency, medical professionals will need to find the appropriate antivenom based on the type of snake venom the patient has been exposed to and administer the antivenom to the patient as soon as possible. However, the time required to transport the patient to the hospital and determine the type of snake venom suffered by the patient, as well as the difficulty in preserving the antivenom serum, are clinical challenges for such emergency treatment. Therefore, those skilled in the art are still committed to developing a new strategy that is more effective against the effects of snake venom.

[0003] Isorhamnetin and quercetin are two flavonoid compounds found in medicinal plants such as Hippophae rhamnoides L. and are known to have anti-tumor and anti-adipogenesis effects and can treat enteritis. However, whether isorhamnetin and quercetin are effective against snake toxins is still unknown. Summary of the invention

[0004] Therefore, an object of the present disclosure is to provide a method for antisnake venom that alleviates at least one disadvantage of the prior art and comprises administering to a subject in need thereof a pharmaceutical composition comprising a flavonoid compound selected from the group consisting of isorhamnetin, quercetin, and combinations thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0005] Other features and advantages of the present disclosure will become apparent in the following detailed description of embodiments with reference to the accompanying drawings. It should be noted that various features may not be drawn to scale.

[0006] Figure 1 The survival rates measured for the pathological control group of Example 1 and experimental groups 1 and 2 are shown below.

[0007] Figure 2 The survival rates of the pathological control group 3 and the experimental group 3 of Example 1 below are shown.

[0008] Figure 3 The survival rates of the pathological control group 4 and the experimental group 4 in Example 1 are shown below.

[0009] Figure 4 The survival rates of the pathological control group 5 and the experimental group 5 in Example 1 below are shown.

[0010] Figure 5 The survival rates of the pathological control group 6 and the experimental group 6 of Example 1 below are shown.

[0011] Figure 6 The survival rates of the pathological control group 7 and the experimental group 7 in Example 1 are shown below.

[0012] Figure 7 The areas of skin ulcers measured in the pathological control group of Example 2 and in each of the experimental groups 1 and 2 are shown below.

[0013] Figure 8 The following shows the areas of skin ulcers measured in the pathological control group 3 and the experimental group 3 of Example 2. DETAILED DESCRIPTION

[0014] It should be appreciated that if any prior art publication is cited herein, this citation does not constitute an admission that the publication forms part of the general knowledge in the art.

[0015] For the purposes of this specification, it will be expressly understood that the term "comprising" means "including but not limited to," and that the term "comprises" has a corresponding meaning.

[0016] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art to which the present disclosure belongs. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used to implement the present disclosure. Of course, the present disclosure is in no way limited to the methods and materials described.

[0017] In developing methods to combat snake envenomation, the applicant unexpectedly discovered that isorhamnetin and quercetin, two flavonoid compounds, can prevent death and alleviate tissue necrosis caused by various snake venoms (of different venomous snakes). On the other hand, the applicant also discovered that apigenin cannot produce such a death-preventing effect, and even accelerates death despite having a structure similar to quercetin and isorhamnetin. Therefore, isorhamnetin and quercetin are expected to be effective in combating snake envenomation.

[0018] Therefore, the present disclosure provides a method for combating the effects of snake venom, comprising administering to an individual in need thereof a pharmaceutical composition comprising a flavonoid compound. The flavonoid compound is selected from the group consisting of isorhamnetin, quercetin, and combinations thereof. In some embodiments, the flavonoid compound may be isorhamnetin. In some embodiments, the flavonoid compound may be quercetin.

[0019] As used herein, the terms "snake venom effect" and "toxicity induced by snake venom" are used interchangeably and refer to various toxicities known to be induced by snake venom, including, but not limited to: neurotoxicity, hemotoxicity, vascular toxicity, cardiotoxicity, myotoxicity, and cytotoxicity.

[0020] In some embodiments, the venom effect can be caused by a hemorrhagic snake venom. In some embodiments, the hemorrhagic snake venom can be from Protobothrops spp. Examples of Protobothrops species may include, but are not limited to: Protobothrops cornutus, Protobothrops selegans, Protobothrops flavoviridis, Protobothrops shimalayanus, Protobothrops jerdonii, Protobothrops kelomohy, Protobothrops mangshanensis, Protobothrops maolanensis, Protobothrops sieversorum, Protobothrops tokarensis, Protobothrops sieversorum ... In some embodiments, the Protobothrops species may be Protobothrops mucrosquamatus.

[0021] In some embodiments, the venom effect can be caused by a neurotoxin. In some embodiments, the neurotoxin can be from a species of the genus Naja (Naja spp.). Examples of species of the genus Naja may include, but are not limited to: Naja annulifera, Naja arabica, Naja ashei, Naja haje, Naja kaouthia, Naja katiensis, Naja mandalayensis, Naja melanoleuca, Naja mossambica, Naja nana, Naja janigricincta, Naja nubiae, Naja oxiana, Naja pallida, Naja philippinensis, Naja sagittifera, Naja samarensis, Naja savannula, Naja senegal, Naja savannula, Naja philippinensis, Naja samarensis, Naja savannula, Naja senegal ... In some embodiments, the Naja spitting cobra can be selected from the group consisting of Naja senegalensis, Naja siamensis, Naja sputatrix, Naja subfulva, Naja sumatrana, Naja nivea, Naja atra, Najanigricolis, and Naja naja. In some embodiments, the Naja spitting cobra can be selected from the group consisting of Naja nivea, Naja atra, Najanigricolis, and Naja naja, and combinations thereof.

[0022] According to the present disclosure, the pharmaceutical composition can be manufactured into a dosage form suitable for parenteral administration, oral administration or topical administration using techniques well known to those skilled in the art.

[0023] According to the present disclosure, the pharmaceutical composition may further include a pharmaceutically acceptable carrier widely used in drug manufacturing technology. For example, the pharmaceutically acceptable carrier may include one or more agents selected from the following: solvents, buffers, emulsifiers, suspending agents, decomposing agents, disintegrants, dispersants, binders, excipients, stabilizers, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption delay agents, liposomes and the like. The selection and quantity of these agents fall within the professional qualities and routine technical scope of those skilled in the art.

[0024] Regarding parenteral administration, the pharmaceutical composition according to the present disclosure can be formulated into an injection using techniques well known to those skilled in the art, such as a sterile aqueous solution or dispersion. In some embodiments, the pharmaceutical composition can be administered by intramuscular injection or subcutaneous injection.

[0025] According to the present disclosure, dosage forms suitable for oral administration include, but are not limited to, sterile powders, lozenges, tablets, buccal lozenges, pills, capsules, dispersible powders or fine granules, solutions, suspensions, emulsions, syrups, elixirs, concentrates and the like.

[0026] According to the present disclosure, the pharmaceutical composition can be manufactured into an external preparation suitable for topical application to the skin using techniques well known to those skilled in the art. The external preparation includes, but is not limited to, emulsions, gels, ointments, creams, patches, liniments, powders, aerosols, sprays, lotions, serums, pastes, foams, drops, suspensions, salves, and bandages.

[0027] According to the present disclosure, the external preparation is prepared by mixing the pharmaceutical composition with a base well known to those skilled in the art.

[0028] According to the present disclosure, the substrate may contain one or more additives selected from the following: water, alcohols, glycols, hydrocarbons [such as petroleum jelly and white petrolatum], waxes [such as paraffin and yellow wax], preservatives, antioxidants, surfactants, absorption enhancers, stabilizing agents, gelling agents [such as 941( 941), microcrystalline cellulose and carboxymethylcellulose], active agents, humectants, odor absorbers, fragrances, pH adjusting agents, chelating agents, emulsifiers, occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants and propellants, etc. The selection and amount of these additives are within the professional qualities and routine technical scope of those skilled in the art.

[0029] As used herein, the terms "administration" and "administering" are used interchangeably and refer to introducing, providing or delivering the pharmaceutical composition to a subject showing symptoms of snake venom effects by any appropriate route to perform its intended effect.

[0030] As used herein, the term "alleviating" or "alleviation" means at least partially reducing, ameliorating, relieving, controlling, treating or eliminating one or more clinical signs of a disease or disorder; as well as lowering, delaying, stopping or reversing the progression of the severity of a condition or symptom being treated and preventing or reducing its likelihood or probability.

[0031] As used herein, the term "subject" means any mammal of interest, such as humans, monkeys, cows, sheep, horses, pigs, goats, dogs, cats, mice, and rats. In some embodiments, the subject is a human.

[0032] The dosage and frequency of administration of the pharmaceutical composition of the present disclosure will vary depending on the severity of the disease (illness) or disorder (disorder) to be treated, the route of administration, and the age, physical condition and response of the individual to be treated. In general, the pharmaceutical composition can be administered in a single dose or in multiple doses.

[0033] The present disclosure will be further described with reference to the following examples, but it should be understood that the following examples are only for illustration and should not be construed as limitations on the implementation of the present disclosure.

[0034] Example

[0035] General experimental materials:

[0036] 1. Experimental mice:

[0037] The Institute of Cancer Research (ICR) mice (5 weeks old, weighing about 10 g-15 g) used in the following experiments were purchased from BioLASCO Co., Ltd. All experimental mice were housed in an animal room under the following laboratory conditions: alternating 12-hour light and 12-hour dark cycles, maintained at a temperature of 24°C, and maintained at a relative humidity of 65%. In addition, all experimental mice were free to drink water and eat. All experimental procedures related to experimental mice were in accordance with the guidelines of the Institutional Animal Care and Use Committee of National Tsing Hua University (Hsinchu, Taiwan, China).

[0038] 2. Flavonoid compound:

[0039] Isorhamnetin, quercetin and apigenin (three flavonoid compounds) used in the following examples were purchased from Sigma-Aldrich.

[0040] Example 1. Evaluation of the efficacy of flavonoid compounds in combating death caused by snake envenomation

[0041] Experimental Materials:

[0042] A.Snake venom

[0043] The venoms from different snake species used in this example and their corresponding median lethal doses (LD50) for ICR mice were 50 ) are shown in Table 1 below.

[0044] Table 1

[0045]

[0046] Experimental methods and results:

[0047] A. The efficacy of flavonoid compounds in combating death caused by hemorrhagic snake venom

[0048] ICR mice were randomly divided into 4 groups, including 1 pathological control group, 2 experimental groups (i.e., experimental groups 1 and 2) and 1 comparison group (n=7 mice in each group). Next, each of isorhamnetin, quercetin and apigenin was mixed with the venom of Agkistrodon acutus at a volume ratio of 1:20, and then incubated at 37°C for 30 minutes to form respective mixed solutions. Then, each ICR mouse in each of the experimental groups 1 and 2 and the comparison group was intradermally injected with an appropriate amount of the mixed solution as shown in Table 2 below. In addition, each ICR mouse in the pathological control group was intradermally injected with the venom of Agkistrodon acutus. The treatment agents and their dosages used for each group are summarized in Table 2 below.

[0049] Table 2

[0050]

[0051] Afterwards, the number of surviving ICR mice in each group was observed and recorded every hour until 10 hours after the treatment agent injection. The survival rate (%) of each group of ICR mice was calculated using the following formula (1):

[0052] A=(B / 7)×100 (1)

[0053] Where: A = survival rate (%)

[0054] B = Number of ICR mice alive per hour until 10 hours after treatment injection

[0055] Reference Figure 1 , the survival rate measured in the pathological control group dropped sharply in the first hour after the treatment agent injection, and continued to drop to 0% in the fifth hour after the treatment agent injection. Furthermore, the survival rate of the ICR mice in the comparison group dropped sharply to 0% in the second hour after the treatment agent injection, while the survival rate of the ICR mice in the experimental group 2 was 25% in the second hour after the treatment agent injection and remained at the same level until the tenth hour after the treatment agent injection. In addition, the survival rate of the ICR mice in the experimental group 1 exceeded 70% in the second hour after the treatment agent injection and even remained above 55% in the tenth hour after the treatment agent injection. These experimental results show that both quercetin and isorhamnetin can effectively fight against death caused by hemorrhagic snake venom, thereby prolonging survival time, while apigenin, although having a structure similar to quercetin and isorhamnetin, not only did not show this effect (that is, fighting death and prolonging survival time), but accelerated death. B. The effectiveness of isorhamnetin in fighting death caused by neurotoxic snake venom

[0056] Based on the results of item A of this example, which show that isorhamnetin has the best effect among other flavonoid compounds in combating the death caused by hemorrhagic snake venom (i.e., the venom of Agkistrodon acutus), the applicant intends to further study the effectiveness of isorhamnetin in combating the death caused by neurotoxic snake venom.

[0057] First, ICR mice were randomly divided into 8 groups, including 4 pathological control groups (i.e., pathological control groups 3 to 6) and 4 experimental groups (i.e., experimental groups 3 to 6) (n=7 mice per group). Next, isorhamnetin was mixed with each of the venom of black-necked cobra, the venom of golden cobra, the venom of Indian cobra, and the venom of Chinese cobra at a volume ratio of 1:20, and then incubated at 37°C for 30 minutes to form respective mixed solutions. Then, each ICR mouse in each of the experimental groups 3 to 6 was intradermally injected with an appropriate amount of the mixed solution as shown in Table 3 below. In addition, each ICR mouse in each of the pathological control groups 3 to 6 was intradermally injected with the corresponding venom of black-necked cobra, the venom of golden cobra, the venom of Indian cobra, and the venom of Chinese cobra. The treatment agents and their dosages for each group are summarized in Table 3 below.

[0058] Table 3

[0059]

[0060] Thereafter, the number of surviving ICR mice in each group was observed and recorded every hour until 14 hours after the treatment agent injection. The survival rate (%) of each group of ICR mice was calculated using formula (1).

[0061] Reference Figure 2 , the survival rate of the pathological control group 3 was reduced to 30% at 4 hours after the injection of the treatment agent, while the survival rate of the experimental group 3 was maintained at 100% until 14 hours after the injection of the treatment agent. Figure 3 The survival rate of the pathological control group 4 was reduced to below 20% at 5 hours after the treatment agent injection and continued to decrease to 0% at 10 hours after the treatment agent injection, while the survival rate of the experimental group 4 was higher than 65% at 10 hours after the treatment agent injection and maintained above 45% at 14 hours after the treatment agent injection. Figure 4 , the survival rate of the pathological control group 5 was reduced to 0% in the first hour after the treatment agent injection, while the survival rate of the experimental group 5 was higher than 65% in the first hour after the treatment agent injection and maintained above 30% in the 14th hour after the treatment agent injection. Figure 5The survival rate of the pathological control group 6 was reduced to 0% at 2 hours after the injection of the treatment agent, while the survival rate of the experimental group 6 was higher than 75% at 2 hours after the injection of the treatment agent and remained above 50% at 14 hours after the injection of the treatment agent.

[0062] These experimental results indicate that isorhamnetin can effectively counteract the death caused by neurotoxic snake venom and thus prolong survival time.

[0063] C. The efficacy of isorhamnetin in alleviating the toxicity caused by neurotoxic snake venom

[0064] To closely simulate the situation where the individual is an emergency patient of a venomous snake bite, in the present application, ICR mice were injected with golden cobra venom and isorhamnetin separately (with a time interval in between) rather than simultaneously (ie, premixed for injection).

[0065] First, ICR mice were randomly divided into 2 groups, including 1 pathological control group (i.e., pathological control group 7) and 1 experimental group (i.e., experimental group 7) (n=7 mice in each group). Then, each ICR mouse in each group was intradermally injected with an appropriate amount of golden cobra venom (dose: 2.5×LD per mouse). 50 ). 30 minutes later, each ICR mouse of the experimental group was intradermally injected with an appropriate amount of isorhamnetin (dose: 50 nmole per mouse).

[0066] Thereafter, the number of surviving ICR mice in each group was observed and recorded every hour until 48 hours after the treatment agent injection. The survival rate (%) of each group of ICR mice was calculated using formula (1).

[0067] Reference Figure 6 , the survival rate of the pathological control group 7 was reduced to 0% at 5 hours after the treatment agent injection, while the survival rate of the experimental group 7 was higher than 40% at the same time and maintained higher than 20% from 12 hours to 28 hours after the treatment agent injection. In addition, some ICR mice in the experimental group 7 can even survive until 48 hours after the treatment agent injection. These experimental results indicate that the administration of isorhamnetin to an individual affected by snake venom can effectively counteract the death caused thereby, thereby prolonging the survival time.

[0068] Example 2. Evaluation of the efficacy of flavonoid compounds in relieving cutaneous ulcers caused by snake venom proteins

[0069] In order to further study the effectiveness of isorhamnetin and quercetin in relieving skin ulcers caused by snake venom proteins, the applicant used a mixture of snake venom proteins containing a molecular weight of not less than 43kD and cardiotoxin that may cause tissue necrosis to conduct experiments to avoid sudden death of ICR mice due to other toxins that may interfere with observation. Experimental materials:

[0070] 1. Preparation of snake venom protein mixture:

[0071] First, cardiotoxin was isolated from an appropriate amount of Chinese cobra venom according to the method described in SC Sue et al., (2001), Biochemistry, 40, 12782-12794. In addition, another appropriate amount of Chinese cobra venom was dissolved in a buffer solution containing 50 mM phosphate and 150 mM sodium chloride (pH 6.4) to form a mixture, and then the mixture was used. The mixture was subjected to gel filtration chromatography using a 75 column (GE Healthcare Life Sciences) to collect high molecular weight proteins with a molecular weight of not less than 43 kD. Then, the cardiotoxin and the high molecular weight protein were mixed to obtain a snake venom protein mixture containing 0.5 mg / mL cardiotoxin and 0.025 mg / mL high molecular weight protein.

[0072] Experimental methods and results:

[0073] A. Effect of flavonoid compounds injected intradermally against skin ulcers caused by snake venom proteins

[0074] First, ICR mice were randomly divided into 3 groups, including 1 pathological control group and 2 experimental groups (i.e., experimental groups 1 and 2) (n=7 mice in each group). Next, each of quercetin and isorhamnetin and an appropriate amount of a mixture of snake venom proteins were mixed at a volume ratio of 1:20, and then incubated at 37°C for 30 minutes to form respective mixed solutions. Then, each ICR mouse of experimental groups 1 and 2 was intradermally injected with an appropriate amount of the mixed solution as shown in Table 4 below. In addition, each ICR mouse of the pathological control group was intradermally injected with a mixture of snake venom proteins. The treatment agents and their dosages for each group are summarized in Table 4 below.

[0075] Table 4

[0076]

[0077] At 24 hours after the treatment agent injection, the skin ulcers developed on the ICR mice in each group were photographed and their areas were analyzed using ImageJ software.

[0078] Reference Figure 7 The areas of skin ulcers measured in Experimental Groups 1 and 2 were significantly reduced compared with those in the pathological control group, among which the area of ​​skin ulcers measured in Experimental Group 2 had the highest reduction.

[0079] B. The effect of topical administration of isorhamnetin on skin ulcers caused by snake venom proteins

[0080] To further evaluate whether isorhamnetin can be administered topically immediately after a subject suffers from a self-venomous snake bite, similar results to those of Experimental Group 2 in Item A of this Example can be obtained, the following experiment was conducted.

[0081] First, 3.1 mg of isorhamnetin was mixed with 1 mL of petroleum jelly in a melted state at 45°C to obtain isorhamnetin ointment. Then, the ICR mice were randomly divided into 2 groups, including 1 pathological control group (i.e., pathological control group 3) and 1 experimental group (i.e., experimental group 3) (n=7 mice in each group). Next, each ICR mouse in each group was intradermally injected with an appropriate amount of snake venom protein mixture (0.1 mL per mouse). Afterwards, an appropriate amount of isorhamnetin ointment was immediately applied to the injection site of each ICR mouse in experimental group 3 (0.2 g / cm 2 , once a day), and as for the ICR mice in the pathological control group 3, the same dose of vaseline was applied to each mouse.

[0082] Reference Figure 8 The area of ​​skin ulcers in the experimental group was significantly reduced compared with that in the pathological control group.

[0083] Furthermore, the applicant further determined the median effective dose (ED50) of isorhamnetin and quercetin for reducing the area of ​​skin ulcers caused by snake venom proteins. 50 ). The results of the test showed that when administered via intradermal injection, the 50% effective dose of isorhamnetin and quercetin was 0.2mM and 1.2mM respectively, and when administered topically, the 50% effective dose of isorhamnetin was 0.6mM. The results showed that isorhamnetin and quercetin can effectively fight against tissue necrosis caused by snake venom proteins through different administration routes.

[0084] In summary, it can be seen that isorhamnetin and quercetin can not only counteract the death caused by snake venom, but also alleviate the tissue necrosis caused thereby, and therefore are expected to be used to counteract the effects of snake venom or snake toxins.

[0085] In the above description, for the purpose of illustration, many specific details have been described for a thorough understanding of the specific example. However, it will be apparent to those skilled in the art that one or more other specific examples can be implemented without some of these specific details. It should also be understood that "one embodiment", "one embodiment", a specific example with a serial number, etc., mentioned throughout this specification, means that a specific feature, structure or characteristic can be included in the implementation of the present disclosure. It should be further understood in the description that, in order to simplify the present disclosure and help understand the purpose of various inventive aspects, various features are sometimes collected in a single specific example, figure or its description, which does not mean that each of these features needs to be implemented in the presence of all other features. In other words, in any of the specific examples described, when the implementation of one or more features or specific details does not affect the implementation of another one or more features or specific details, the one or more features can be selected and implemented separately without the other one or more features or specific details. It should be further noted that in implementing the present disclosure, one or more features or details from one embodiment may be implemented together with one or more features or details from another embodiment, where appropriate.

[0086] While the present disclosure has been described with reference to specific examples which are considered to be exemplary, it is to be understood that the disclosure is not limited to the specific examples disclosed, but is intended to cover a variety of different arrangements to be included in the spirit and scope of the broadest interpretation to include all such modifications and equivalent arrangements.

Claims

1. A method for combating the effects of snake venom, comprising administering to a subject in need thereof a pharmaceutical composition comprising a flavonoid compound selected from the group consisting of isorhamnetin, quercetin, and combinations thereof.

2. The method of claim 1, wherein the envenomation is caused by a hemorrhagic snake venom.

3. The method of claim 2, wherein the hemorrhagic snake venom is from a species of the genus Agkistrodon.

4. The method of claim 3, wherein the Agkistrodon species is Agkistrodon acutus.

5. The method of claim 1, wherein the venom effect is caused by a neurotoxic snake venom.

6. The method of claim 5, wherein the neurotoxic snake venom is from a species of the genus Naja.

7. The method of claim 6, wherein the species of the genus Naja is selected from the group consisting of: golden cobra, Chinese cobra, black-necked cobra, Indian cobra, and combinations thereof.

8. The method according to any one of claims 1 to 7, wherein the pharmaceutical composition is in a dosage form for parenteral administration, oral administration or topical administration.

9. A pharmaceutical composition for use in combating the effects of snake venom, the use of which comprises administering the pharmaceutical composition to an individual in need thereof, the pharmaceutical composition comprising a flavonoid compound selected from the group consisting of isorhamnetin, quercetin, and combinations thereof.

10. The pharmaceutical composition for use in combating the effect of snake venom according to claim 9, wherein the effect of snake venom is caused by hemorrhagic snake venom.

11. The pharmaceutical composition for use in combating the effects of snake venom according to claim 10, wherein the hemorrhagic snake venom is from a species of the genus Agkistrodon.

12. The pharmaceutical composition for use in combating the effects of snake venom according to claim 11, wherein the species of the genus Agkistrodon is Agkistrodon acutus.

13. The pharmaceutical composition for use in combating the effect of snake venom according to claim 9, wherein the effect of snake venom is caused by a neurotoxin.

14. The pharmaceutical composition for use in combating the effects of snake venom according to claim 13, wherein the neurotoxic snake venom is from a species of the genus Naja.

15. The pharmaceutical composition for use in combating the effects of snake venom according to claim 14, wherein the species of the genus Naja is selected from the group consisting of: golden cobra, Chinese cobra, black-necked cobra, Indian cobra, and combinations thereof.

16. The pharmaceutical composition for use against the effects of snake venom according to any one of claims 9 to 15, wherein the pharmaceutical composition is in a dosage form for parenteral administration, oral administration or topical administration.

17. A composition for use in the preparation of a medicament for counteracting the effects of snake venom in a subject, wherein the composition comprises a flavonoid compound selected from the group consisting of isorhamnetin, quercetin, and combinations thereof.

18. The use according to claim 17, wherein the snake venom effect is caused by hemorrhagic snake venom.

19. The use according to claim 18, wherein the hemorrhagic snake venom is from a species of the genus Agkistrodon.

20. The use according to claim 19, wherein the species of the genus Agkistrodon is Agkistrodon acutus.

21. The use according to claim 17, wherein the venom effect is caused by a neurotoxic snake venom.

22. The use according to claim 21, wherein the neurotoxic snake venom is from a species of the genus Naja.

23. The use according to claim 22, wherein the species of the genus Naja is selected from the group consisting of: golden cobra, Chinese cobra, black-necked cobra, Indian cobra, and combinations thereof.

24. The use according to any one of claims 17 to 23, wherein the composition is in a dosage form for parenteral administration, oral administration or topical administration.

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