Composition containing coenzyme Q10 and application thereof in heart

Abstract

The invention relates to the field of biological medicine, in particular to a coenzyme Q10-containing composition and application thereof in the heart, and the coenzyme Q10-containing composition comprises a coenzyme Q10 sustained-release system, perilla oil, gelatin, glycerin and water; the coenzyme Q10 sustained-release system comprises a self-assembled polypeptide as shown in SEQ ID NO: 17. On the basis of earlier-stage research, the self-assembled polypeptide is further improved, and finally the self-assembled polypeptide shown as SEQ ID NO: 17 is screened out, the self-assembled polypeptide has higher encapsulation efficiency and drug loading capacity and can better deliver drugs, and animal experiments prove that the self-assembled polypeptide can better treat myocardial injury and protect the heart, so that the self-assembled polypeptide has good application prospects. The method has important clinical value and economic value.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a composition containing coenzyme Q10 and application thereof in the heart. Background Art

[0002] Coenzyme Q10 is a fat-soluble molecule produced by various tissues of the human body. It consists of a central benzoquinone part with a 10-unit polyisoprene lipid tail attached to it. In mitochondria, Coenzyme Q10 plays a key role as a mitochondrial electron carrier in the process of oxidative phosphorylation and participates in the formation of mitochondrial ATP. At the same time, Coenzyme Q10 has redox properties, which can affect the formation of reactive oxygen species, and acts as an antioxidant to prevent oxidative damage caused by reactive oxygen species, reduce and neutralize free radicals (including reactive oxygen species). In addition, Coenzyme Q10 is speculated to exert anti-inflammatory effects by reducing nuclear transcription factor κB (NF-κB). NF-κB is activated by reactive oxygen species, which in turn upregulates the expression of proinflammatory cytokines (such as tumor necrosis factor, interleukin 6, etc.). As an antioxidant, Coenzyme Q10 can neutralize free radicals and inhibit the activation of NF-κB, thereby inhibiting the expression of proinflammatory cytokines. Supplementation of Coenzyme Q10 can promote the production of ATP, alleviate the reduction of ATP production caused by mitochondrial dysfunction, and effectively improve the energy metabolism disorder of cardiomyocytes. At the same time, in the respiratory chain, coenzyme Q10 transfers electrons from complex I or complex II to complex III, reducing the generation of reactive oxygen species, which helps to alleviate the impact of increased reactive oxygen generation on mitochondrial function. At the same time, coenzyme Q10 has antioxidant function, which can remove free radicals, reduce oxidative stress damage, and improve myocardial damage.

[0003] Therefore, coenzyme Q10 has the functions of protecting the heart, controlling heart diseases, and strengthening the myocardium. It is also found that once the coenzyme Q10 in the myocardial cells decreases, myocardial ischemia will occur. Supplementing coenzyme Q10 can enhance the blood supply to the myocardial cells, resist heart failure, and resist arrhythmia.

[0004] For example, the prior art CN 117180245 B discloses a coenzyme Q10 composition and its preparation process and application in protecting the heart. The drug is a multi-layer sustained-release drug prepared by stacking a degradable isolation layer and a drug layer layer by layer. Compared with traditional sustained-release drugs, the preparation process of the sustained-release drug is simpler and more efficient, and the sustained-release effect is also very good. It can effectively control the release rate of coenzyme Q10 in the patient's body and achieve a more efficient therapeutic effect.

[0005] Self-assembling peptides (SAP) can trigger the intramolecular free energy spontaneously or under certain conditions, driving the molecules to self-assemble into nanostructures with higher physical and chemical stability. The internal driving force that triggers the self-assembly of peptide molecules is mainly non-covalent forces, such as electrostatic interactions, hydrogen bonds, л-л stacking, and hydrophilic / hydrophobic interactions. SAP has a rich number of modifiable functional groups, good biocompatibility, high drug loading, long drug retention time, and precise targeted recognition sites. In addition, SAP has flexible response capabilities and can respond to local microenvironmental stimuli of the lesion, such as pH, temperature, light, ionic strength, and enzyme catalysis, and has been widely used in tissue engineering, anti-tumor, and antibacterial applications.

[0006] For example, the prior art CN 116869971 B discloses a coenzyme Q10 sustained-release soft capsule and a preparation method. The soft capsule contains a coenzyme Q10 sustained-release system, perilla oil, gelatin, glycerol and water. The preparation method of the coenzyme Q10 sustained-release system is as follows: 1) dissolving the self-assembling polypeptide FFGFAFIGRHHH in water; 2) ultrasonically dissolving coenzyme Q10 in an ethanol solution; 3) slowly adding ethanol dissolved with coenzyme Q10 dropwise to the self-assembling polypeptide aqueous solution, then evaporating the ethanol, centrifuging, and lyophilizing the supernatant to obtain a sample lyophilized powder. Under the optimal parameter conditions, the drug loading of the self-assembling polypeptide carrying coenzyme Q10 is 14%, and the encapsulation rate is 42%; the coenzyme Q10 sustained-release system capsule can effectively prolong the half-life of coenzyme Q10.

[0007] However, the sequence of the self-assembling polypeptides in the above-mentioned prior art is single, and the drug loading capacity and encapsulation efficiency need to be improved. Therefore, it is urgent to explore more self-assembling polypeptides and compositions containing coenzyme Q10 with better drug loading capacity and encapsulation efficiency. Summary of the invention

[0008] In order to solve the above problems, the present invention first provides a composition containing coenzyme Q10, which comprises a coenzyme Q10 sustained-release system, perilla oil, gelatin, glycerol and water; the coenzyme Q10 sustained-release system comprises a self-assembling polypeptide as shown in SEQ ID NO: 17.

[0009] In certain embodiments, the method for preparing the sustained-release system of coenzyme Q10 comprises the following steps:

[0010] 1) Dissolve the self-assembling polypeptide in water;

[0011] 2) dissolving coenzyme Q10 in ethanol solution by ultrasonication;

[0012] 3) Slowly add ethanol containing coenzyme Q10 dropwise into the self-assembling polypeptide aqueous solution, then evaporate the ethanol, centrifuge, and freeze-dry the supernatant to obtain a sample freeze-dried powder.

[0013] In certain embodiments, the coenzyme Q10 sustained-release system comprises 40-50 parts, perilla oil 10-15 parts, gelatin 20-30 parts, glycerol 10-20 parts, and water 1-10 parts, by weight.

[0014] In certain embodiments, the coenzyme Q10 sustained-release system comprises 45 parts, perilla oil 12 parts, gelatin 28 parts, glycerol 10 parts, and water 5 parts.

[0015] In certain embodiments, the concentration of the self-assembling polypeptide is 1-2 g / L.

[0016] In certain embodiments, the ultrasound power is 200-350W, and the ultrasound time is 5-10min.

[0017] In certain embodiments, the centrifugal process parameter is 2500 r / min for 12 min.

[0018] The present invention also provides a method for preparing the above-mentioned composition containing coenzyme Q10, comprising the following steps:

[0019] 1) dissolving the self-assembling polypeptide in water; dissolving coenzyme Q10 in an ethanol solution by ultrasonication; slowly adding the ethanol containing coenzyme Q10 dropwise into the aqueous solution of the self-assembling polypeptide, then evaporating the ethanol, centrifuging, and lyophilizing the supernatant to obtain a sample lyophilized powder;

[0020] 2) preparing contents by mixing the freeze-dried powder and perilla oil according to a certain proportion;

[0021] 3) preparing soft capsules by mixing gelatin and glycerin according to a certain proportion;

[0022] 4) Packing the contents and soft capsules into soft capsules containing a sustained release system.

[0023] Finally, the present invention provides an application of the above-mentioned composition containing coenzyme Q10, characterized in that the application is for preparing a drug for treating heart-related diseases.

[0024] In certain embodiments, the heart-related disease is myocardial damage.

[0025] Compared with the prior art, the present invention has at least the following beneficial effects:

[0026] Based on previous research, the present invention further improved the self-assembling polypeptide and finally screened out the self-assembling polypeptide as shown in SEQ ID NO: 17, which has a higher encapsulation rate and drug loading capacity, can better deliver drugs, and animal experiments have shown that it can better treat myocardial injury and protect the heart, and has great clinical and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Pharmacokinetic comparison. DETAILED DESCRIPTION

[0028] In order to make the technical problems, technical solutions and advantages to be solved by the present invention more clear, a detailed description will be given below with reference to the accompanying drawings and specific embodiments.

[0029] Example 1 Synthesis of self-assembling polypeptides

[0030] Referring to the applicant's prior authorized document CN116869971B, single-point and multi-point mutants were designed based on FFGFAFIGRHHH (SEQ ID NO: 1). The results are shown in Table 1, and the underlined amino acids are mutation sites.

[0031] Table 1 Mutants of self-assembling polypeptides

[0032]

[0033]

[0034] The self-assembling polypeptide of Table 1 was synthesized by solid phase synthesis and purified by HPLC to obtain the self-assembling polypeptide of Table 1 with a purity of 95%.

[0035] Example 2 Preparation of soft capsules

[0036] Preparation of soft capsules:

[0037] 1) Dissolve the self-assembling polypeptide of Table 1 in water at a concentration of 1.5 g / L; dissolve coenzyme Q10 in ethanol solution, and ultrasonicate for 8 min at an ultrasonic power of 300 W; slowly add ethanol containing coenzyme Q10 dropwise to the self-assembling polypeptide aqueous solution, with a ratio of ethanol to water of 1:10, then evaporate the ethanol, centrifuge at 2500 r / min for 12 min, and freeze-dry the supernatant to obtain a sample freeze-dried powder;

[0038] 2) preparing contents by mixing the freeze-dried powder and perilla oil in a ratio of 45:12;

[0039] 3) preparing soft capsules by mixing gelatin, glycerin and water in a ratio of 28:10:5;

[0040] 4) Packing the contents and soft capsules into soft capsules containing a sustained release system.

[0041] Determination of drug loading and encapsulation efficiency of the composition containing coenzyme Q10 using each self-assembling polypeptide:

[0042] Drug loading (%) = dosage of coenzyme Q10 - concentration measured in liquid phase × solution volume / total amount of lyophilized powder of sustained-release system loaded with coenzyme Q10;

[0043] Encapsulation efficiency (%) = dosage of coenzyme Q10 - concentration measured in liquid phase × volume of solution / dosage of coenzyme Q10.

[0044] The results are shown in Table 2.

[0045] Table 2 Drug loading and encapsulation efficiency of each group

[0046] Peptide mutant number Drug loading (%) Encapsulation rate (%) SEQ ID NO: 1 12.0 42.1 SEQ ID NO: 2 14.0 45.3 SEQ ID NO: 3 13.0 45.0 SEQ ID NO: 4 12.4 42.5 SEQ ID NO: 5 11.7 41.3 SEQ ID NO: 6 12.6 43.4 SEQ ID NO: 7 15.6 46.7 SEQ ID NO: 8 14.2 45.6 SEQ ID NO: 9 16.3 49.3 SEQ ID NO: 10 16.4 49.3 SEQ ID NO: 11 14.7 45.9 SEQ ID NO: 12 17.1 49.6 SEQ ID NO: 13 16.3 49.3 SEQ ID NO: 14 18.1 49.9 SEQ ID NO: 15 15.7 47.1 SEQ ID NO: 16 18.7 50.3 SEQ ID NO: 17 23.6 56.3

[0047] From the results in Table 2, it can be seen that the drug loading (%) and encapsulation efficiency (%) of the self-assembling polypeptide shown in SEQ ID NO: 17 were significantly improved (P<0.01). Therefore, the subsequent experiments were carried out using the polypeptide of SEQ ID NO: 17.

[0048] Comparative Example 1

[0049] Preparation of soft capsules:

[0050] 1) Coenzyme Q10, water and perilla oil were mixed in a ratio of 18.9:26.1:12 to prepare the contents;

[0051] 2) preparing soft capsules by mixing gelatin, glycerol and water in a ratio of 28:10:5;

[0052] 3) Packing the contents and soft capsules into soft capsules containing a sustained release system.

[0053] Example 3 Sustained Release Effect Test of Each Soft Capsule

[0054] 18 SD rats weighing about 250g were purchased and divided into 3 groups. The rats were given drugs by oral gavage, and the content of coenzyme Q10 was 50mg / kg. On the second day, coenzyme Q10, soft capsules of comparative example 1, and soft capsules of example 2 (SEQ ID NO: 17) were given at the same time. Blood samples were taken from the orbital vein of the rats at 0.5, 1, 2, 4, 6, 8, and 10 hours, anticoagulants were added to the blood samples, centrifuged, and the supernatant was taken and stored at 4°C.

[0055] Take a sample and use a high performance liquid chromatograph to test the concentration of coenzyme Q10. Figure 1 As shown, the composition capsule containing Coenzyme Q10 prepared using the self-assembled polypeptide of SEQ ID NO: 17 can effectively prolong the half-life of Coenzyme Q10.

[0056] Example 4

[0057] Several SD rats weighing about 200g were selected, and myocardial ischemic injury model rats were prepared by coronary artery clamping method. The rats with successful modeling were selected and evenly divided into blank group, control group and treatment group, with 10 rats in each group, wherein the blank group was not treated with any drugs during the monitoring period, the control group was administered 0.2mg / kg coenzyme Q10 every day, and the treatment group (SEQID NO: 17) was administered 0.2mg / kg coenzyme Q10 sustained-release drug every day, and the drug was administered for one week. After the drug administration was completed, the range of myocardial infarction after coronary artery ligation and the content of SOD and MDA in serum were counted. The results are shown in Table 3 below. The composition containing coenzyme Q10 prepared using the self-assembling polypeptide of SEQ ID NO: 17 can significantly reduce the area of ​​myocardial infarction, has a good protective and repair effect on myocardial ischemic injury, and can increase the activity of SOD in rat serum after sustained-release drug treatment, inhibit the generation of MDA, and has a protective effect on myocardial ischemia (P<0.05).

[0058] Table 3. Protective effect of the composition containing coenzyme Q10 on myocardial ischemia

[0059] Group Myocardial infarction area (%) SOD (NU / ml) MDA (nmol / ml) Blank Group 18.41±1.22 198.50±9.41 9.11±0.81 Control group 14.55±1.31 223.94±7.99 6.31±0.52 Treatment Group 7.24±1.24 260.11±10.11 4.21±0.54

[0060] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composition containing coenzyme Q10, characterized in that: The invention comprises a coenzyme Q10 sustained-release system, perilla oil, gelatin, glycerol and water; the coenzyme Q10 sustained-release system comprises a self-assembling polypeptide as shown in SEQ ID NO:

17.

2. The composition containing coenzyme Q10 according to claim 1, characterized in that The preparation method of the coenzyme Q10 sustained-release system comprises the following steps: 1) Dissolve the self-assembling polypeptide in water; 2) dissolving coenzyme Q10 in ethanol solution by ultrasonication; 3) Slowly add ethanol containing coenzyme Q10 dropwise into the self-assembling polypeptide aqueous solution, then evaporate the ethanol, centrifuge, and freeze-dry the supernatant to obtain a sample freeze-dried powder.

3. The composition containing coenzyme Q10 according to claim 1, characterized in that By weight, the coenzyme Q10 sustained-release system comprises 40-50 parts, perilla oil 10-15 parts, gelatin 20-30 parts, glycerol 10-20 parts, and water 1-10 parts.

4. The composition containing coenzyme Q10 according to claim 1, characterized in that 45 parts of coenzyme Q10 sustained-release system, 12 parts of perilla oil, 28 parts of gelatin, 10 parts of glycerol, and 5 parts of water.

5. The composition containing coenzyme Q10 according to claim 1, characterized in that The concentration of the self-assembling polypeptide is 1-2 g / L.

6. The composition containing coenzyme Q10 according to claim 1, characterized in that: The ultrasonic power is 200-350W, and the ultrasonic time is 5-10min.

7. The composition containing coenzyme Q10 according to claim 1, characterized in that The centrifugal process parameters are 2500r / min for 12min.

8. A method for preparing a composition containing coenzyme Q10 as claimed in any one of claims 1 to 7, comprising the following steps: 1) dissolving the self-assembling polypeptide in water; dissolving coenzyme Q10 in an ethanol solution by ultrasonication; slowly adding the ethanol containing coenzyme Q10 dropwise into the aqueous solution of the self-assembling polypeptide, then evaporating the ethanol, centrifuging, and lyophilizing the supernatant to obtain a sample lyophilized powder; 2) preparing contents by mixing the freeze-dried powder and perilla oil according to a certain proportion; 3) preparing soft capsules by mixing gelatin and glycerin according to a certain proportion; 4) Packing the contents and soft capsules into soft capsules containing a sustained release system.

9. Use of a composition containing coenzyme Q10 as claimed in any one of claims 1 to 7, characterized in that: The application is to prepare medicines for treating heart-related diseases.

10. The use according to claim 9, characterized in that: The heart-related disease is myocardial damage.

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