Coenzyme Q10-containing composition and application thereof in treatment of heart-related disease

Abstract

Coenzyme Q10-containing composition comprising perilla oil, gelatin, glycerin, water and a coenzyme Q10 (CoQ10) sustained-release system comprising a self-assembling peptide consisting of Seq ID No: 1

Description

TECHNICAL FIELD

[0001] This application relates to biological medicine, and more particularly to a coenzyme QlO-containing composition, and an application thereof in the treatment of a heart-related disease. BACKGROUND

[0002] Coenzyme Q10 is a liposoluble molecule produced by various tissues in human body, consisting of a central benzoquinone ring with a 10-unit polyisoprene lipid tail attached thereto. In mitochondria, coenzyme Q10 plays a crucial role as an electron carrier in the process of oxidative phosphorylation to participate in the formation of mitochondrial adenosine triphosphate (ATP). Moreover, owing to the redox properties, Coenzyme Q10 can affect the formation of reactive oxygen species (ROS) and act as an antioxidant to prevent oxidative damage caused by ROS, thereby reducing and neutralizing free radicals (including ROS). In addition, considering the fact that NF-kB can be activated by ROS to up-regulate the expression of proinflammatory cytokines (such as tumor necrosis factor and interleukin-6, and coenzyme Q10 can act as an antioxidant to neutralize free radicals and inhibit the activation of NF-kB to suppress the expression of pro-inflammatory cytokines, coenzyme Q10 is speculated to exhibit anti-inflammatory activity by reducing the level of nuclear transcription factor kB(NF-kB). Supplementation of coenzyme Q10 can promote the production of ATP to relieve the decline in ATP generation caused by mitochondrial dysfunction and effectively improve the energy metabolism disorder in myocardial cells. Meanwhile, in the respiratory chain, coenzyme Q10 can transfer electrons from complex I or complex II to complex III, reducing the production of ROS and mitigating the adverse effects of ROS on the mitochondrial function. Coenzyme Q10 also has an antioxidant activity to scavenge the free radicals, relieve oxidative stress injury and ameliorate myocardial damage.

[0003] Therefore, coenzyme Q10 plays a role in protecting the heart, controlling heart-related diseases and strengthening the myocardial function. Once coenzyme Q10 in myocardial cells decreases, myocardial ischemia will occur. Supplementation with coenzyme Q10 can enhance the blood supply to myocardial cells, and prevent heart failure and arrhythmia.

[0004] Chinese patent No. 117180245 B provides a coenzyme QlO-containing composition, and its preparation and application in protecting the heart. This drug has a multilayer sustained-release structure prepared by layer-by-layer stacking of degradable isolation layers and drug layers. Compared with traditional sustained-release drugs, this preparation process is simpler and more efficient, also demonstrating excellent sustained-release performance. It can effectively control the release rate of coenzyme Q10 in the patient's body, thereby achieving a more efficient therapeutic effect.

[0005] Self-assembling peptide (SAP) can trigger intramolecular free energy either spontaneously or under certain specific conditions, driving the molecules to selfassemble into nanostructures with higher physical and chemical stability. The internal driving forces that trigger the self-assembly of peptide molecules are mainly non-covalent interactions, such as electrostatic interactions, hydrogen bonding, 7t-7t stacking, and hydrophilic / hydrophobic interactions. SAPs offer abundant modifiable functional groups, and has good biocompatibility, high drug-loading capacity, prolonged drug retention and precise targeting recognition. In addition, SAPs exhibit a flexible response capability to stimuli in the local lesion microenvironmental, such as pH, temperature, light, ionic strength and enzymatic activity. It has been widely applied in tissue engineering, anti-tumor and antibacterial fields.

[0006] Chinese patent No. 116869971 B provides a coenzyme Q10 sustained-release soft capsule and a preparation method thereof. The soft capsule contains a coenzyme Q10 sustained-release system, perilla oil, gelatin, glycerin and water. The preparation method of the coenzyme Q10 sustained-release system is performed as follows:

[0007] (1) a self-assembling peptide FFGFAFIGRHHH is dissolved in water;

[0008] (2) coenzyme Q10 is ultrasonically dissolved in an ethanol solution; and

[0009] (3) the coenzyme Q10 ethanol solution is dropwise added to the aqueous solution of the self-assembling peptide; then the system is subjected to evaporation to remove ethanol and centrifugation, and a supernatant is collected, and freeze-dried to give the desired product in the form of freeze-dried powder.

[0010] Under the optimal conditions, the drug-loading capacity of coenzyme Q10 on the self-assembling peptide is 14%, and the encapsulation rate is 42%. The coenzyme Q10 sustained-release capsules can effectively prolong the half-life of coenzyme Q10.

[0011] However, the self-assembling peptides in the above-mentioned existing technologies exhibit limited sequence diversity, and the drug-loading capacity and the encapsulation rate still need to be improved. Therefore, it is urgently needed to develop more self-assembling peptides and coenzyme QlO-containing compositions having better drug-loading capacity and encapsulation rate. SUMMARY

[0012] In order to solve the above problems, the present disclosure provides a coenzyme QlO-containing composition, comprising a coenzyme Q10 sustained-release system, perilla oil, gelatin, glycerin and water, wherein the coenzyme Q10 sustained-release system comprises a self-assembling peptide (SAP) consisting of SEQ ID NO: 17.

[0013] In some embodiments, the coenzyme Q10 sustained-release system is prepared through steps of:

[0014] (1) a self-assembling peptide is dissolved in water to obtain a peptide solution;

[0015] (2) coenzyme Q10 is ultrasonically dissolved in an ethanol solution to obtain a coenzyme Q10 solution; and

[0016] (3) the coenzyme Q10 solution is dropwise added to the peptide solution to obtain a mixed system; then the mixed system is subjected to evaporation for ethanol removal and centrifugation; and a supernatant is collected, and freeze-dried to give coenzyme Q10 sustained-release system in a form of freeze-dried powder.

[0017] In some embodiments, the coenzyme QlO-containing composition comprises 40-50 parts by weight of the coenzyme Q10 sustained-release system, 10-15 parts by 3 weight of the perilla oil, 20-30 parts by weight of gelatin, 10-20 parts by weight of glycerin and 1-10 parts by weight of water.

[0018] In some embodiments, the coenzyme QlO-containing composition comprises 45 parts by weight of the coenzyme Q10 sustained-release system; 12 parts by weight of the perilla oil; 28 parts by weight of the gelatin; 10 parts by weight of the glycerin; and 5 parts by weight of the water.

[0019] In some embodiments, a concentration of the self-assembling peptide in the peptide solution is 1-2 g / L.

[0020] In some embodiments, an ultrasonic power is 200-350 W, and an ultrasonic time is 5-10 min.

[0021] In some embodiments, the centrifugation is performed at 2500 rpm for 12 min.

[0022] The present disclosure provides the method of preparing the coenzyme QlO-containing composition comprising:

[0023] (1) the self-assembling peptide is dissolved in water to obtain a peptide solution; coenzyme Q10 is ultrasonically dissolved in an ethanol solution to obtain a coenzyme Q10 solution; the coenzyme Q10 solution is dropwise added to the peptide solution, followed by evaporation for ethanol removal and centrifugation; and a supernatant is collected, and freeze-dried to give the coenzyme Q10 sustained-release system in a form of freeze-dried powder;

[0024] (2) the coenzyme Q10 sustained-release system is mixed with the perilla oil according to a preset weight ratio to prepare a core material;

[0025] (3) the gelatin is mixed with the glycerin according to a preset weight ratio to prepare a soft capsule shell; and

[0026] (4) the core material is encapsulated into the soft capsule shell to prepare the coenzyme QlO-containing composition in a form of soft capsule.

[0027] The present disclosure also provides an application of the coenzyme QlO-containing composition in the preparation of a drug for treating a heart-related disease.

[0028] In some embodiments, the heart-related disease is myocardial damage.

[0029] Compared with the prior art, the present disclosure has the following beneficial effects.

[0030] Based on the prior researches, the present disclosure further structurally improves the self-assembling peptides, and screens the self-assembling peptide consisting of SEQ ID NO: 17, which has a higher encapsulation rate and drug-loading capacity, and can more effectively deliver the active drug component. Moreover, it has been demonstrated by animal experiments that the composition of the present disclosure can effectively treat the myocardial injury and protect the heart, and has a significant clinical and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Fig. 1 shows comparison between example 2 and comparative example 1 in terms of pharmacokinetics. DETAILED DESCRIPTION OF EMBODIMENTS

[0032] To make the to-be-solved technical problems, technical solutions and advantages of the present disclosure clearer, the disclosure will be described in detail below in combination with the accompanying drawings and specific embodiments. EXAMPLE 1

[0033] Reference could be made to the applicant’s previous Chinese patent No. 116869971B for the synthesis of the self-assembling peptides. Based on the sequence FFGFAFIGRHHH (SEQ ID NO: 1), single-point and multi-point mutants were designed, and the results were shown in Table 1, with the underlined amino acid residue representing the mutation site.

[0034] Table 1 Self-assembling peptide mutants Peptide mutant No. Sequence SEQ ID NO: 1 FFGFAFIGRHHH SEQ ID NO: 2 FFRFAFIGRHHH SEQ ID NO: 3 FFGQAFIGRHHH SEQ ID NO: 4 FFGFCFIGRHHH SEQ ID NO: 5 FFGFASIGRHHH SEQ ID NO: 6 FFGFAFMGRHHH SEQ ID NO: 7 FFGFAFIKRHHH SEQ ID NO: 8 FFGFAFIGGHHH SEQ ID NO: 9 FFGFAFIGRNHH SEQ ID NO: 10 FFGFAFIGQHHH SEQ ID NO: 11 FFGRAFIERHHH SEQ ID NO: 12 FFGFGFIGRVHH SEQ ID NO: 13 FFHFAGIGRHHH SEQ ID NO: 14 FFGQAFQGRHHH SEQ ID NO: 15 FFGFEFITRHHH SEQ ID NO: 16 FFFFAQIGMHHH SEQ ID NO: 17 FFGAAFRGRAHH

[0035] The self-assembling peptides listed in Table 1 were synthesized by solid-phase synthesis and purified by HPLC to reach a purity of 95% or more. EXAMPLE 2 Preparation of soft capsule

[0036] (1) The self-assembling peptides listed in Table 1 were respectively dissolved in water to produce a series of peptide solutions with a concentration of 1.5 g / L. Coenzyme Q10 was ultrasonically dissolved in an ethanol solution with an ultrasonic power of 300 W and an ultrasonic time of 8 min to obtain a coenzyme Q10 solution. The coenzyme Q10 solution was dropwise added to the peptide solutions with a ratio of 1: 10 (ethanol: water) to obtain a mixed system. Then, the mixed system was subjected to evaporation for ethanol removal and centrifugation at 2500 rpm for 12 min. A supernatant was freeze-dried to give the coenzyme Q10 sustained-release system in a form of freeze-dried.

[0037] (2) The freeze-dried powder was mixed with the perilla oil in a weight ratio of 45: 12 to prepare a core material;

[0038] (3) The gelatin, the glycerin and the water were mixed in a weight ratio of 28: 10: 5 to prepare the soft capsule shell.

[0039] (4) The core material was encapsulated into the soft capsule shell to prepare the coenzyme QlO-containing composition in a form of soft capsule.

[0040] The drug-loading capacity and the encapsulation rate of the coenzyme QlO-containing compositions prepared with individual self-assembling peptides were determined respectively according to the following formulas:

[0041] drug-loading capacity (%) = (dosage of coenzyme Q10 - concentration measured in liquid phase * solution volume) / total amount of freeze-dried powder of sustained-release system loaded with coenzyme Q10; and

[0042] entrapment rate (%) = (dosage of coenzyme Q10 - concentration measured in liquid phase * solution volume) / dosage of coenzyme Q10.

[0043] The results were shown in Table 2.

[0044] Table 2 Drug-loading capacity and encapsulation rate of each group Peptide mutant No. Drug-loading capacity (%) 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

[0045] It could be observed from Table 2 that the self-assembling peptide represented by SEQ ID NO: 17 demonstrated significantly improved drug-loading capacity (%) and encapsulation rate (%) (P<0.01). Therefore, the subsequent experiments were conducted with the self-assembling peptide of SEQ ID NO: 17. COMPARATIVE EXAMPLE 1

[0046] Preparation of the soft capsule

[0047] (1) The coenzyme Q10, water and perilla oil were mixed in a weight ratio of 18.9: 26.1: 12 to prepare the core material.

[0048] (2) Gelatin, glycerin and water were mixed in a weight ratio of 28: 10: 5 to prepare the soft capsule shell.

[0049] (3) The core material was encapsulated into the soft capsule shell to produce the sustained-release system-containing soft capsule. EXAMPLE 3 Test of sustained-release performance

[0050] Eighteen SD (Sprague-Dawley) rats, weighing approximately 250 g, were purchased and divided into three groups. On the 2nd day, the three groups were intragastrically administered with 50 mg / kg of coenzyme Q10, the soft capsule of comparative example 1 and the soft capsule of Example 2 (SEQ ID NO: 17) at the same time, respectively.

[0051] Blood samples were collected from orbital veins of individual rats at 0.5, 1, 2, 4, 6, 8, and 10 h post the administration, added with an anticoagulant and centrifuged. The supernatants were collected and stored at 4°C.

[0052] The samples were analyzed by high-performance liquid chromatography (HPLC) to determine the concentration of coenzyme Q10. As shown in Fig. 1, the coenzyme QlO-containing composition capsule with the self-assembling peptide represented by SEQ ID NO: 17 could effectively prolong the half-life of coenzyme Q10. EXAMPLE 4

[0053] Several SD (Sprague-Dawley) rats, weighing approximately 200 g, were selected, and treated by coronary artery ligation to establish myocardial ischemia injury models. Successfully modeled rats were evenly divided into a blank group, a control group, and a treatment group, with 10 rats in each group. The blank group received no drug treatment during an observation period; the control group was administered with 0.2 mg / kg of coenzyme Q10 daily; and the treatment group (SEQ ID NO: 17) was administered with 0.2 mg / kg of the sustained-release coenzyme Q10 drug daily.

[0054] The administration was performed consecutively for one week. After the administration, the myocardial infarction area after the coronary artery ligation and serum SOD (superoxide dismutase) and MDA (Malondialdehyde) levels were determined. As shown in Table 3, the coenzyme QlO-containing composition with the self-assembling peptide of SEQ ID NO: 17 significantly reduced the myocardial infarction area, exhibiting good protective and repairing effects on the myocardial ischemic injury. Additionally, the treatment with the sustained-release drug increased the serum SOD activity, inhibited the MDA production, and exhibited a protective effect on the myocardial ischemia (P <0.05).

[0055] Table 3 Protective effect of the coenzyme QlO-containing composition on the myocardial ischemia 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

[0056] Described above are merely preferred embodiments of the present disclosure, which are not intended to limit the present disclosure. It should be understood that various improvements and modifications made by those skilled in the art without departing from the spirit of the present disclosure shall fall within the scope of the present disclosure defined by the appended claims.

Claims

enzyme Q10-containing composition, comprising:a coenzyme Q10 sustained-release system;perilla oil;gelatin;glycerin; andwater;characterized in that the coenzyme Q10 sustained-release system comprises a self-assembling peptide consisting of SEQ ID NO: 17.

2. The coenzyme Q10-containing composition according to claim 1, characterized in that the coenzyme Q10 sustained-release system is prepared through steps of:(1) dissolving the self-assembling peptide in water to obtain a peptide solution;(2) ultrasonically dissolving coenzyme Q10 is in an ethanol solution to obtain a coenzyme Q10 solution; and(3) dropwise adding the coenzyme Q10 solution to the peptide solution to obtain a mixed system; subjecting the mixed system to evaporation for ethanol removal and centrifugation; and collecting and freeze-drying a supernatant to give the coenzyme Q10 sustained-release system in a form of freeze-dried powder.

3. The coenzyme Q10-containing composition according to claim 1, characterized in that the coenzyme Q10-containing composition comprises:40-50 parts by weight of the coenzyme Q10 sustained-release system;10-15 parts by weight of the perilla oil;20-30 parts by weight of the gelatin;10-20 parts by weight of the glycerin; and1-10 parts by weight of the water.

4. The coenzyme Q10-containing composition according to claim 1, characterized in that the coenzyme Q10-containing composition comprises:45 parts by weight of the coenzyme Q10 sustained-release system;12 parts by weight of the perilla oil;28 parts by weight of the gelatin;10 parts by weight of the glycerin; and5 parts by weight of the water.

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

6. The coenzyme Q10-containing composition according to claim 2, characterized in that an ultrasonic power is 200-350 W, and an ultrasonic time is 5-10 min.

7. The coenzyme Q10-containing composition according to claim 2, characterized in that the centrifugation is performed at 2500 rpm for 12 min.

8. A method of preparing the coenzyme Q10-containing composition according to any one of claims 1-7, comprising:(1) dissolving the self-assembling peptide in water to obtain a peptide solution; ultrasonically dissolving coenzyme Q10 in an ethanol solution to obtain a coenzyme Q10 solution; dropwise adding the coenzyme Q10 solution to the peptide solution, followed by evaporation for ethanol removal and centrifugation; and collecting and freeze-drying a supernatant to give the coenzyme Q10 sustained-release system in a form of freeze-dried powder;(2) mixing the coenzyme Q10 sustained-release system with the perilla oil according to a preset weight ratio to prepare a core material;(3) mixing the gelatin with the glycerin according to a preset weight ratio to prepare a soft capsule shell; and(4) encapsulating the core material into the soft capsule shell to prepare the9. The coenzyme Q10-containing composition according to any one of claims 1-7 for use in treating myocardial damage.T +44(0)30 0300 2000Search report under Section 17 of the Patents Act 1977Application No.: GB2514819.8Claims searched: 1-10Date search completed: 3 March 2026International classificationSubclass and subgroup Valid from A61K31 / 122 01 / 01 / 2006 A61K47 / 42 01 / 01 / 2017 A61P9 / 00 01 / 01 / 2006 A61P9 / 10 01 / 01 / 2006 C07K7 / 08 01 / 01 / 2006Field of searchWorldwide search of patent documents classified in the following areas of the IPC: A61K, C07KDatabases used in the preparation of this search report:CAS ONLINE; INTERNET; SEARCH-NPL; SEARCH-PATENTDocuments considered to be relevantPatent literatureCategory Relevant to claims Document of relevance A - CN 116869971 A (GUANGDONG RUNHE BIOLOGICAL TECHNOLOGY CO) -paragraph 6-19 A - JP 6429372 B2 (UNIV OF MIYAZAKI) - paragraph 12, 18, 24 Non-patent literature Category Relevant to claims Document of relevanceT +44(0)30 0300 2000Category Relevant to claims Document of relevance A - Food Hydrocolloids, Vol. 72, 13 / 06 / 2017, B. Muhoza et al., "Time effect on coenzyme Q10 loading and stability of micelles based on glycosylated casein via Maillard reaction”, pages 271-280, https: / / doi.org / 10.1016Zj.foodhyd.2017.05.046, see abstractCategoriesLetter or symbol Description X Document indicating lack of novelty or inventive step. Y Document indicating lack of inventive step, if combined with another document of the same category. & Member of the same patent family. A Document indicating technological background. P Document published on or after the priority date but before the filing date of the present application. E Earlier application published on or after the filing date of the present application.

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