Use of pharmaceutical composition in treatment of migraine

Abstract

The present invention relates to a pharmaceutical composition. The pharmaceutical composition comprises cobra cardiotoxin (CDX) and a pharmaceutically acceptable carrier thereof. The pharmaceutical composition is further characterized by comprising use of cobra cardiotoxin (CDX) in combination with any one or both of the following drugs for treating migraine: cobra phospholipase A2 (PLA2) and cobra neurotoxin (NTX). The pharmaceutical composition can be used for preventing and treating migraine and down-regulating the concentration of nitric oxide (NO) in migraine patients. The pharmaceutical composition is of great significance for the treatment of migraine.

Description

Use of a pharmaceutical composition for the treatment of migraine TECHNICAL FIELD

[0001] The present invention relates to the use of a composition for the manufacture of a medicament for the treatment of migraine in a patient. BACKGROUND

[0002] Migraine is a common, recurrent headache disorder characterized by severe throbbing pain on one or both sides of the head, often accompanied by nausea, vomiting, hypersensitivity to light and sound, and other symptoms. Common types include migraine without aura, migraine with aura, and chronic migraine. The causes of migraine are likely related to genetics, endocrine metabolism, environmental factors, mental factors, and other factors, and often have a genetic background. Female patients are 2-3 times more likely than males, and most patients are in adolescence, while a small number of patients may be in childhood.

[0003] Symptomatically, the typical presentation of migraine is severe headache, which can be unilateral headache, which can migrate to the other side, and can also be sensitive to light, noise, and odor. Common associated symptoms include nausea, vomiting, stomach discomfort, abdominal pain, feeling very hot or cold, pale complexion, and other symptoms.

[0004] In terms of treatment, migraine cannot be cured at present, and most treatments aim to reduce headache attacks, relieve associated symptoms, and prevent headache recurrence. There are drug treatment and non-drug treatment, and drug treatment is divided into attack period treatment and preventive treatment. Clinical drugs include opioid analgesics, non-steroidal analgesics, and monoclonal antibodies, but they cannot produce satisfactory therapeutic effects, and some drugs have very serious side effects, and frequent use can cause other clinical symptoms in patients. Therefore, the current migraine treatment drugs have not been able to truly meet the clinical needs of patients, and migraine has been a disease that seriously affects the quality of human life.

[0005] The causes of migraine are generally believed to be genetic, endocrine metabolism, environmental factors, mental factors, and other factors. Further research has found that migraine is associated with pathological factors such as increased serum nitric oxide (NO) concentration, because increased serum nitric oxide (NO) concentration can cause dilation of the brain blood vessels, thereby causing migraine, and nitric oxide can cause pain hypersensitivity and inflammatory response, which can exacerbate the degree of migraine, [1-5] and reducing nitric oxide (NO) levels is an effective method for improving the degree of migraine.

[0006] Current migraine treatment options include acute and preventive treatment. With the exploration of migraine attack-related signaling pathways, more and more treatment methods will be developed for migraine treatment, and down-regulating nitric oxide (NO) concentration can be used as an effective method for treating migraine. SUMMARY

[0007] The present application relates to a kind of compositions in the preparation of treating patient migraine drug, it is characterized in that, the composition includes cobra heart poison (CDX), the drug composition of the present application has analgesic effect to adopt rat nitroglycerin type migraine test model.Its mechanism can be related to blocking or down-regulating nitric oxide (NO) signal path, cobra heart poison can simultaneously treat and prevent the occurrence of migraine, and there is no serious toxic side effect in animal experiment, so it is a desirable drug candidate.

[0008] Cobra heart poison has the commonality of three-finger protein structure in functional structure, they not only same spatial structure, and the amino acid sequence of their mature protein has high homology, so the commonality of heart poison in functional structure makes them all achieve consistent regulation nitric oxide (NO) signal path function, which also makes heart poison in the common consistency of treating rat nitroglycerin type migraine model.

[0009] The amino acid sequence of the mature protein of cobra heart poison has the following common characteristics:

[0010] lkcnklxpx xxktcxagkn lcykmfmxxx xxxpvkrgci dvcpkxsxlv kyvccntdrc n (SEQ ID No.1)

[0011] In some embodiments, the amino acid sequence of cobra heart poison in the composition is

[0012] lkcnklvpl fyktcpagkn lcykmfmvat pkvpvkrgci dvcpkssllv kyvccntdrc n (SEQ ID No2)

[0013] In some embodiments, the amino acid sequence of cobra heart poison in the composition is

[0014] lkcnklipi asktctagkn lcykmfmmsd ltipvkrgci dvcpknsllv kyvccntdrc n (SEQ ID No3)

[0015] In some embodiments, the amino acid sequence of cobra heart poison in the composition is

[0016] lkcnklipi asktcpagkn lcykmfmmsd ltipvkrgci dvcpknsllv kyvccntdrc n (SEQ ID No4)

[0017] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0018] lkcnklipi asktcpagkn lcykmfmvat pkvpvkrgci dvcpknsllv kyvccntdrc n (SEQ ID No 5)

[0019] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0020] lkcnklvpl fyktcpagkn lcykmfmvat pkvpvkrgci dvcpknsalv kyvccntdrc n (SEQ ID No 6)

[0021] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0022] lkcnklvpl fyktcpagkn lcykmfmvsn kmvpvkrgci dvcpknsalv kyvccntdrc n (SEQ ID No 7)

[0023] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0024] lkcnklvpl fyktcpagkn lcykmfmvsn ltvpvkrgci dvcpknsalv kyvccntdrc n (SEQ ID No 8)

[0025] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0026] lkcnklipi asktcpagkn lcykmfmvat pkvpvkrgci dvcpkssllv kyvccntdrc n (SEQ ID No 9)

[0027] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0028] lkcnklipia sktcpagknl cykmfmmsdl tipvkrgcid vcpknsllvk yvccntdrcn (SEQ ID No 10)

[0029] In some embodiments, the amino acid sequence of cobrotoxin in the composition is

[0030] lkcnklipla yktcpagknl cykmfmvsnk tvpvkrgcid acpknsllvk yvccntdrcn (SEQ ID No 11)

[0031] In some embodiments, the amino acid sequence of the cobrotoxin in the composition is

[0032] lkcnklipla yktcpagknl cykmfmvsnk tvpvkrgcid acpknsllvk yvccntdrcn (SEQ ID No 11)

[0033] In some embodiments, the amino acid sequence of the cobrotoxin in the composition is

[0034] lkcnklipla yktcpagknl cykmfmvsnk tvpvkrgcid acpknsllvk yvccntdrcn (SEQ ID No 11)

[0035] In some embodiments, the amino acid sequence of the cobrotoxin in the composition is

[0036] lkcnklipla yktcpagknl cykmfmvsnk tvpvkrgcid acpknsllvk yvccntdrcn (SEQ ID No 11)

[0037] In some embodiments, the composition is administered once daily for 3-120 consecutive days.

[0038] In some embodiments, the composition is administered once every 12 hours, twice daily, for 3-120 consecutive days.

[0039] In some embodiments, the composition is administered once every 8 hours, three times daily, for 3-120 consecutive days.

[0040] In some embodiments, the cobrotoxin in the composition is administered at a dose of 0.01-100 micrograms per kilogram per administration.

[0041] In some embodiments, the cobrotoxin in the composition is administered at a dose of 80 micrograms per kilogram per administration.

[0042] In some embodiments, the cobrotoxin in the composition is administered at a dose of 60 micrograms per kilogram per administration.

[0043] In some embodiments, the dose of cobra heart toxin administered in the composition is 40 micrograms per kilogram per administration.

[0044] In some embodiments, the dose of cobra heart toxin administered in the composition is 20 micrograms per kilogram per administration.

[0045] In some embodiments, the dose of cobra heart toxin administered in the composition is 10 micrograms per kilogram per administration.

[0046] In some embodiments, the dose of cobra heart toxin administered in the composition is 5.0 micrograms per kilogram per administration.

[0047] In some embodiments, the dose of cobra heart toxin administered in the composition is 2.5 micrograms per kilogram per administration.

[0048] In some embodiments, the dose of cobra heart toxin administered in the composition is 1.0 micrograms per kilogram per administration.

[0049] In some embodiments, the dose of cobra heart toxin administered in the composition is 0.5 micrograms per kilogram per administration.

[0050] Our animal model experiments also found that the combined use of cobra heart toxin (CDX) with either cobra phospholipase A2 (PLA2) or with cobra neurotoxin (NTX) had better analgesic effect on the nitroglycerin-induced migraine test model in rats; and the combined use of cobra heart toxin (CDX) with both cobra phospholipase A2 (PLA2) and cobra neurotoxin (NTX) achieved the strongest analgesic effect, although any one of cobra heart toxin (CDX), cobra phospholipase A2 (PLA2), or cobra neurotoxin (NTX) alone had analgesic effect on migraine.

[0051] The amino acid sequence (FASTA) of cobra phospholipase A2 mature protein used in combination with cobra heart toxin (CDX) or in combination with cobra heart toxin (CDX) and cobra neurotoxin (NTX) has the following common features:

[0052] nly qfknmiqctv psrswwdfad ygcycgkggs gtpvddldrc cqvhdxcyxe aekisgcwpy xktysyecsq gtltckggnn

[0053] acaaavcdcd rlaaicfaga pyxxxxynix lkarcq (SEQ ID No. 15)

[0054] Further, in some combination use embodiments, the following four amino acid sequences of phospholipase A2 showed more effective effects on treating the rat nitroglycerin type migraine model.

[0055] In some combination use embodiments, the amino acid sequence of phospholipase A2 in the composition is

[0056] nly qfknmiqctv psrswwdfad ygcycgrggs gtpvddldrc cqvhdncyne aekisgcwpy fktysyecsq gtltckggnn acaaavcdcd rlaaicfaga pynnnnynid lkarcq (SEQ ID No. 16)

[0057] In some combination use embodiments, the amino acid sequence of phospholipase A2 in the composition is

[0058] nly qfknmiqctv psrswwdfad ygcycgrggs gtpvddldrc cqvhdhcyne aekisgcwpy sktysyecsq gtltckggnn

[0059] acaaavcdcd rlaaicfaga pynnnnynid lkarcq (SEQ ID No. 17)

[0060] In some combination use embodiments, the amino acid sequence of phospholipase A2 in the composition is

[0061] nly qfknmvqctv pnrswwdfad ygcycgrggs gtpvddldrc cqvhdncyge aekisrcwpy fktysyecsq gtltckggnn

[0062] acaaavcdcd rlaaicfaga pyndnnynid lkarcq (SEQ ID No. 18)

[0063] In some combination use embodiments, the amino acid sequence of phospholipase A2 in the composition is

[0064] nly qfknmiqctv psrswwdfad ygcycgrggs gtpvddldrc cqvhdncyne aekisgcwpy fktysyecsq gtltckg

[0065] gnnacaaavc dcdrlaaicf agapyndndy ninlkarc (SEQ ID No. 19)

[0066] The amino acid sequences of the mature protein of the cobra neurotoxin (NTX) in combination with cobra cardiotoxin (CDX), or in simultaneous combination with cobra cardiotoxin (CDX) and cobra phospholipase A2 (PLA2) (FASTA) have the following common features:

[0067] lechnqqsxq tptttgcsgg etncykkrwr dhrgyrterg cgcpxvkngi einccttdrc nn (SEQ ID No. 20)

[0068] Further, in some combination embodiments, the following three amino acid sequences of the cobra neurotoxin show more effective effects on treating the rat nitroglycerin migraine model.

[0069] lechnqqssq tptttgcsgg etncykkrwr dhrgyrterg cgcpsvkngi einccttdrc nn (SEQ ID No. 21)

[0070] lechnqqssq tptttgcsgg etncykkrwr dhrgyrterg cgcpivkngi esnccttdrc nn (SEQ ID No. 22)

[0071] lechnqqsiq tptttgcsgg etncykkrwr dhrgyrterg cgcpsvkngi einccttdrc nn (SEQ ID No. 23)

[0072] In some combination embodiments, the amino acid sequence of the mature protein of the cobra neurotoxin in the composition is SEQ ID No. 21

[0073] In some combination embodiments, the amino acid sequence of the mature protein of the cobra neurotoxin in the composition is SEQ ID No. 22

[0074] In some combination embodiments, the amino acid sequence of the mature protein of the cobra neurotoxin in the composition is SEQ ID No. 23

[0075] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0076] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0077] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0078] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0079] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0080] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0081] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0082] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0083] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0084] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0085] In some combination embodiments, the weight ratio of cobra cardiotoxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any ratio from 0.01% to 100% cobra cardiotoxin (CDX) to 99.9% to 0% cobra phospholipase A2 (PLA2).

[0086] In some combination embodiments, the combination of cobra cardiotoxin (CDX), cobra phospholipase A2 (PLA2), and cobra neurotoxin (NTX) in the composition is in a weight ratio of 0.01-100% cobra cardiotoxin (CDX) to 50-0% cobra neurotoxin (NTX) to 49.9-0% phospholipase A2 (PLA2).

[0087] In some combination embodiments, the combination of cobra cardiotoxin, neurotoxin, and phospholipase A2 in the composition is in a weight ratio of 0.01-60%: 50-30%: 49.9-10%

[0088] In some combination embodiments, the combination of cobra cardiotoxin, neurotoxin, and phospholipase A2 in the composition is in a weight ratio of 0.01-59%: 50-29%: 49.9-12%

[0089] In some combination embodiments, the combination of cobra cardiotoxin, neurotoxin, and phospholipase A2 in the composition is in a weight ratio of 0.01-58%: 50-28%: 49.9-14%

[0090] In some combination embodiments, the combination of cobra cardiotoxin, neurotoxin, and phospholipase A2 in the composition is in a weight ratio of 0.01-54%: 50-30%: 49.9-16%

[0091] In some embodiments, the cobra cardiotoxin, cobra phospholipase A2, and cobra neurotoxin can be extracted from snake venom or obtained by recombinant or synthetic techniques.

[0092] In some embodiments, the dosage form of the composition is a sublingual film.

[0093] In some embodiments, the dosage form of the composition is an oral dosage form.

[0094] In some embodiments, the dosage form of the composition is an injectable dosage form.

[0095] In some embodiments, the dosage form of the composition is a nasal spray dosage form.

[0096] In some embodiments, the dosage form of the composition is a transdermal dosage form.

[0097] In some embodiments, the pharmaceutically acceptable excipient is selected from one or more of a protein stabilizer, a film excipient, a mucosa penetration enhancer, a cosolvent, a solvent, and a preservative.

[0098] In some embodiments, the protein stabilizer is mannitol.

[0099] In some embodiments, the film excipient is propylene glycol, polyethylene glycol, hydroxypropyl beta cyclodextrin, tween 80, hydroxypropyl methyl cellulose (HPMC), methyl cellulose, xanthan gum.

[0100] In some embodiments, the mucosa permeation enhancer is laurocapram, poloxamer, bornyl alcohol, dextrobornyl alcohol.

[0101] In some embodiments, the co-solvent is propylene glycol.

[0102] In some embodiments, the solvent agent is sodium chloride, ammonium acetate.

[0103] In some embodiments, the preservative is benzyl alcohol, benzalkonium chloride.

[0104] Another advantage of the present application is in production, because the cobra heart toxin, phospholipase A2, and neurotoxin disclosed in the present application have a clear amino acid sequence, so they can be produced by genetic engineering, solving the practical problem of scarcity of snake venom resources; even if the heart toxin is obtained by continuing to separate and purify natural snake venom, it is easier to control the quality and purity because of the clear amino acid sequence in the process, which lays a necessary foundation for the development of pharmaceuticals of monomer components in snake venom.

[0105] The present application is further described below in conjunction with specific examples, but the following examples are not a limitation of the present application; at the same time, any equivalent replacement in the art according to the disclosure of the present application shall fall within the scope of protection of the present application. DETAILED DESCRIPTION EXAMPLE

[0106] Example 1: Obtaining of cobra heart toxin protein (SEQ ID No. 2)

[0107] Dissolve 1 g of cobra crude venom in 25 ml of 0.025 mol / L ammonium acetate buffer at pH 6.0, centrifuge at low temperature, and take the supernatant; equilibrate the TSK CM-650 (M) column with 0.025 mol / L ammonium acetate solution at pH 6.0; after loading, perform 2-step gradient elution (0.1~0.5 mol and 0.6~1 mol) with 0.1~0.5 mol and 0.7~1.0 mol ammonium acetate buffer at pH 5.9, UV detection parameters: 280 nm; elution flow rate: 48 ml / h; collect various toxin components according to the recorded spectrum, and collect multiple protein peaks in the collected solution; perform N-terminal sequencing on each protein peak, and further perform Edman degradation sequencing on the protein with N-terminal lkcnkl, and finally obtain various sequence proteins of cobra heart toxin.

[0108] The amino acid sequence of the primary structure of cobra heart toxin (SEQ ID No. 2) in Fasta form is: lkcnklvplfyktcpagknlcykmfmvatpkvpvkrgcidvcpkssllvkyvccntdrcn

[0109] Example 2: Analgesic effect of cobra heart toxin (CDX); or cobra heart toxin (CDX) combined with cobra phospholipase A2 (PLA2); or cobra heart toxin (CDX) combined with cobra phospholipase A2 (PLA2) and cobra neurotoxin (NTX) on experimental migraine model rats of nitroglycerin type

[0110] 1. Grouping of experimental animals and modeling

[0111] 1.1 Grouping

[0112] Take 80 SD rats with a body weight of 180-220 g, and randomly divide them into 10 normal control rats, 10 model control rats, 10 CDX treatment rats, 10 CDX prevention rats, 10 CDX+PLA2 treatment rats, 10 CDX+PLA2 prevention rats, 10 CDX+PLA2+NTX treatment rats, and 10 CDX+PLA2+NTX prevention rats, a total of 8 groups.

[0113] 1.2 Modeling

[0114] Modeling method: Except for the normal control group, the rest of the groups were intraperitoneally injected with nitroglycerin 10 mg / kg for 3 consecutive days.

[0115] The CDX prevention group was given nasal drops of 80 μg / kg per time, 2 times / day for 7 consecutive days before modeling to the day of modeling; the CDX+PLA2 prevention group was given nasal drops of 80 μg / kg CDX+20 μg / kg PLA2 per time, 2 times / day for 7 consecutive days before modeling to the day of modeling; and the CDX+PLA2+NTX prevention group was given nasal drops of 60 μg / kg CDX+30 μg / kg NTX+10 μg / kg PLA2 per time, 2 times / day for 7 consecutive days before modeling to the day of modeling.

[0116] When modeling, in addition to the normal control group physiological saline injection, the rest of the groups were injected with nitroglycerin 10 mg / kg, 3 days in a row, as described above. The CDX treatment group, CDX+PLA2 treatment group, and CDX+PLA2+NTX treatment group were administered drugs during the modeling stage. The CDX treatment group was administered 80 g / kg per time, 2 times / day. The CDX+PLA2 treatment group was administered 80 g / kg CDX+20 g / kg PLA2 per time, 2 times / day. The CDX+PLA2+NTX treatment group was administered 60 g / kg CDX+30 g / kg NTX+10 g / kg PLA2 per time, 2 times / day.

[0117] Cobra heart toxin (CDX) SEQ ID No. 2, phospholipase A2 (PLA2) SEQ ID No. 16, and neurotoxin (NTX) SEQ ID No. 21 were used for animal experiments.

[0118] 2. Observation index and detection method

[0119] 2.1 Observation index

[0120] Number of times of scratching head of rats in each time period: After the last modeling, the animal behavior index was observed for 30-120 minutes. From the last modeling, each 30 minutes was taken as a time period, and the number of times of scratching head of rats in each time period was observed by using the continuous time segmentation counting method. (shown in Table 1)

[0121] 2.2 Detection method

[0122] Detection of serum nitric oxide (NO) level: After the behavior experiment detection was completed, the normal group, model control group, CDX treatment group, and CDX prevention group of mice were anesthetized, and blood was taken around the orbits. After standing, the supernatant was obtained by centrifugation at 3000 r / minute for 20 minutes at 4 ℃, and was stored in a-80 ℃ refrigerator. The detection was performed according to the ELISA kit instruction manual. (shown in Table 2)

[0123] Experimental results

[0124] Table 1 Number of times of scratching head of rats

[0125] (x̅±SD, n=10)

[0126] Group 30 (min) 60 (min) 90 (min) 120 (min) Normal group 1.03 ± 0.40 0.83 ± 0.37 0.89 ± 0.53 0.77 ± 0.47 Model control group 30.8 ± 2.82 43.9 ± 5.36 37.6 ± 4.72 24.5 ± 4.10 Prevention group (CDX) 27.2 ± 3.68* 39.6 ± 2.01* 33.8 ± 2.09* 21.1 ± 2.21* Treatment group (CDX) 28.6 ± 6.15* 38.5 ± 4.27* 32.6 ± 4.60* 19.5 ± 3.26* Prevention group (CDX + PLA2) 26.6 ± 3.52* 38.1 ± 3.91* 33.1 ± 4.01* 20.7 ± 3.19* Treatment group (CDX + PLA2) 27.8 ± 3.34* 37.6 ± 2.11** 31.1 ± 4.41** 15.9 ± 3.61** Prevention group (CDX + PLA2 + NTX) 25.1 ± 3.22** 37.2 ± 2.25** 31.8 ± 3.87** 18.8 ± 3.03** Treatment group (CDX + PLA2 + NTX) 25.3 ± 2.98** 35.9 ± 2.63** 29.5 ± 3.14** 14.2 ± 4.01**

[0127] * indicates that the CDX treatment group, CDX prevention group, CDX + PLA2 treatment group and CDX + PLA2 prevention group have significant difference compared with the model control group (P < 0.05.); ** indicates that the CDX treatment group, CDX prevention group, CDX + PLA2 treatment group, CDX + PLA2 prevention group, CDX + NTX + PLA2 treatment group, CDX + NTX + PLA2 prevention group have significant difference compared with the model control group (P < 0.01).

[0128] Table 2 Comparison of nitric oxide (NO) content in blood of normal group, model control group, CDX treatment group and CDX prevention group rats

[0129] (μmol / ml x̅±SD n=10)

[0130] Group NO (μmol / ml) Normal group 27.9 ± 3.44 Model control group 40.3 ± 2.86 CDX prevention group 34.3 ± 4.29** CDX treatment group 31.3 ± 4.9**

[0131] ** indicates that the CDX treatment group and CDX prevention group have significant difference compared with the model control group (P < 0.01.)

[0132] Reference:

[0133] 1. Xu XH, et al. Detection of serum 5⁃HT, NO, MMP⁃9 and CGRP in patients with vestibular migraine and its clinical significance. J Mol Diagn Ther. March 2022, Vol. 14 No. 3

[0134] 2. Liu B, Ma YS, Mao WJ, et al. Experimental study on the changes of plasma nitric oxide and calcitonin gene-related peptide in migraine model rats and the intervention effect of flunarizine. [J]. Shaanxi Journal of Medicine, 2020, 49(7):5⁃7.

[0135] 3. Blaine Jacobs et al;, Neurovascular contributions to migraine: moving beyond vasodilation. Neuroscience. 2016 Dec 3; 338: 130⁃144.

[0136] 4. L Neeb et al;, Nitric oxide in migraine. CNS Neurol Disord Drug Targets. 2007 Aug;6(4):258⁃64.

[0137] 5. Xiao YM. Clinical study on zhennangning capsule combined with sodium ferulate in the treatment of migraine. [J]. Modern Medicine and Clinic, 2019, 34(1):41⁃44.

Claims

1. Use of a composition in the manufacture of a medicament for treating and / or preventing migraine, the composition comprising a therapeutically effective amount of cobra heart toxin (CDX) and pharmaceutically acceptable excipients.

2. Use according to claim (1), characterized in that, The medicament has at least one of the following (1) - (3) uses: (1) the medicament is used for treating migraine; (2) the medicament is used for preventing migraine; (3) the medicament is used for down-regulating the concentration of nitric oxide (NO) in the body of a migraine sufferer.

3. Use according to claim (1), characterized in that, The mature protein of cobra heart toxin (CDX) in the composition has an amino acid sequence shown in SEQ ID No. 1 - SEQ ID No. 14, or a mature protein having 90% or more homology with the amino acid sequence shown in SEQ ID No. 1 - SEQ ID No. 14, respectively, and the function of the mature protein is the same or similar to the function of the mature protein shown in SEQ ID No. 1 - SEQ ID No.

14.

4. The use according to claim (1), further characterized by, The composition further comprises the use of cobra heart toxin (CDX) in combination with any one or both of cobra phospholipase A2 (PLA2) and cobra neurotoxin (NTX) for treating migraine.

5. The composition of claim (4), wherein, The weight ratio of cobra heart toxin (CDX) to cobra phospholipase A2 (PLA2) in the composition is any weight ratio of cobra heart toxin (CDX) 0.01% - 100% to cobra phospholipase A2 (PLA2) 99.9% - 0%; the weight ratio of cobra heart toxin (CDX) to cobra neurotoxin (NTX) in the composition is any weight ratio of cobra heart toxin (CDX) 0.01% - 100% to cobra neurotoxin (NTX) 99.9% - 0%; or the weight ratio of cobra heart toxin (CDX) to cobra phospholipase A2 (PLA2) and cobra neurotoxin (NTX) in the composition is any weight ratio of cobra heart toxin (CDX) 0.01% - 100% to cobra neurotoxin (NTX) 50% - 0% and any weight ratio of cobra phospholipase A2 (PLA2) 49.9% - 0%.

6. The composition of claim (4), further characterized by, The mature protein of the cobra phospholipase A2 in the composition has the amino acid sequence shown in SEQ ID No. 15 - SEQ ID No. 19, or a mature protein having 90% or more homology with the mature protein of the amino acid sequence shown in SEQ ID No. 15 - SEQ ID No. 19, respectively, which has the same or similar function as the mature protein shown in SEQ ID No. 1 - SEQ ID No. 5; the mature protein of the cobra neurotoxin (NTX) in the composition has the amino acid sequence shown in SEQ ID No. 20 - SEQ ID No. 23, or a mature protein having 95% or more homology with the mature protein of the amino acid sequence shown in SEQ ID No. 20 - SEQ ID No. 23, respectively, which has the same or similar function as the mature protein shown in SEQ ID No. 20 - SEQ ID No.

23.

7. The use according to claim (1), characterized in that, The dosage form of the composition is an oral administration dosage form, a sublingual administration dosage form, a nasal administration dosage form, a transdermal administration dosage form, or an injection dosage form.

8. The use according to claim (1), characterized in that, The pharmaceutically acceptable adjuvant in the pharmaceutical composition is selected from one or more of a protein stabilizer, a membrane excipient, a mucosa penetration enhancer, a cosolvent, a solvent, and a preservative.

9. The use according to claim (1), further characterized in that the protein stabilizer in the pharmaceutically acceptable adjuvant is mannitol, the membrane excipient is propylene glycol, polyethylene glycol, hydroxypropyl β-cyclodextrin, Tween-80, hydroxypropyl methylcellulose (HPMC), methylcellulose, xanthan gum, the mucosa penetration enhancer is laurocapram, poloxamer, camphor, dextrocamphor, the cosolvent is propylene glycol, the solvent is sodium chloride, ammonium acetate, and the preservative is benzyl alcohol, benzalkonium chloride.

10. The use according to claim (1), characterized in that, The dosage of the cobra cardiotoxin in the composition comprises from 0.01 μg / kg to 100 μg / kg per time, administered 1-3 times per day.