Anti-vascular aging composition based on Malabaricone C and application thereof

Abstract

The invention provides an anti-vascular aging composition based on Malabaricone C and application thereof, and relates to the technical field of biology, and the anti-vascular aging composition based on Malabaricone C comprises the following components: Malabaricone C and optional auxiliary materials. In the composition, the weight percentage of the Malabaricone C is 0.01%-0.5%, and the weight percentage of the Malabaricone C is 0.01%-0.5%. And the amount concentration of an effective substance of the Malabaricone C for improving senescence of vascular endothelial cells is 0.5-20 [mu] M. The composition can be applied to the field of food or health care products, not only can effectively scavenge free radicals and relieve oxidative stress, but also can improve the vascular endothelial function and inhibit inflammatory response, thereby delaying the blood vessel aging process.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to an anti-vascular aging composition based on Malabaricone C and its application. Background Art

[0002] The main characteristics of vascular aging include the decline of vascular endothelial cell function, the remodeling of the vascular wall structure, and the dysfunction of vascular vasomotor function. These pathological changes are considered to be important driving factors in the occurrence and development of cardiovascular diseases.

[0003] At present, the preventive and intervention measures for vascular aging mainly focus on drug treatment and lifestyle adjustment. However, the existing treatment methods still have many deficiencies, such as limited clinical efficacy and significant potential side effects. This has prompted the academic and industrial circles to urgently seek more efficient and safe new strategies.

[0004] In nature, many plant extracts have been proven to have biological activities such as anti-aging and antioxidant effects. Among them, it has been shown that the compound Malabaricone C is a component extracted from the spice nutmeg, which has anti-inflammatory effects and exerts its anti-inflammatory effect by regulating cellular redox reactions. In addition, Malabaricone C is also considered to be a sphingomyelin synthase inhibitor and can resist diet-induced obesity in mice. We have identified through computer high-throughput screening that Malabaricone C can inhibit the aging of vascular endothelial cells, increase the release of endothelial cell NO, and significantly delay the vascular aging of mice. However, the specific effects and mechanisms of its anti-vascular endothelial cell aging and vascular aging have not been fully studied.

[0005] In view of this, the present invention is specifically proposed. Summary of the Invention

[0006] One of the purposes of the present invention is to provide an application of Malabaricone C in the preparation of anti-vascular aging products, so as to solve at least one of the technical problems existing in the prior art.

[0007] Another purpose of the present invention is to provide an anti-vascular aging composition based on Malabaricone C. The present invention proposes a composition with Malabaricone C as the core, which further enhances its anti-vascular aging effect through synergistic action with other natural active ingredients. This composition can be applied to the fields of food or health products, and can not only effectively scavenge free radicals, reduce oxidative stress, but also improve vascular endothelial function and inhibit inflammatory reactions, thereby delaying the process of vascular aging.

[0008] A third object of the present invention is to provide an application of an anti-vascular aging composition in the preparation of food or health products.

[0009] A fourth object of the present invention is to provide an application of an anti-vascular aging composition in the preparation of anti-vascular aging drugs.

[0010] In order to achieve the above objects of the present invention, the following technical solutions are specifically adopted:

[0011] In a first aspect, the present invention provides an application of Malabaricone C in the preparation of a product for anti-vascular aging.

[0012] In an optional embodiment, the effective substance concentration of Malabaricone C for improving the senescence of vascular endothelial cells is 0.5 - 20 μM.

[0013] In an optional embodiment, the effective concentration of Malabaricone C for improving the senescence of vascular endothelial cells is 10 μM.

[0014] In an optional embodiment, Malabaricone C is used to improve the senescence of vascular endothelial cells, and the vascular endothelial cells include primary human umbilical vein endothelial cells.

[0015] In a second aspect, the present invention provides an anti-vascular aging composition based on Malabaricone C, and its components include Malabaricone C and optional excipients.

[0016] In an optional embodiment, the anti-vascular aging composition includes the following components by weight percentage:

[0017] 0.01% - 0.5% Malabaricone C and the balance of excipients.

[0018] In an optional embodiment, the excipients include one or more of antioxidants and anti-inflammatory agents.

[0019] In an optional embodiment, the oxidants include one or more of vitamin C and catechins;

[0020] The anti-inflammatory agent includes tea polyphenols.

[0021] In a third aspect, the present invention provides an application of an anti-vascular aging composition in the preparation of food or health products.

[0022] In a fourth aspect, the present invention provides an application of an anti-vascular aging composition in the preparation of anti-vascular aging drugs.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The anti-vascular aging composition based on Malabaricone C provided by the present invention, wherein Malabaricone C is used to improve the replicative senescence of vascular endothelial cells and the pathological endothelial cell senescence induced by high glucose, oxidative stress, and 5-fluorouracil chemotherapy drugs, and its specific manifestations are as follows: Malabaricone C can significantly reduce the expression of cell cycle inhibitors P16 and P21 in vascular endothelial cells; the Malabaricone C can significantly reduce the presence of galactosidase staining deposits in endothelial cells;

[0025] The Malabaricone C composition provided by the present invention is used to improve the vascular aging induced by the natural aging model. Its specific manifestations are as follows: the Malabaricone C composition can enhance the NO expression level in the serum of senescent mice; the Malabaricone C composition reduces the levels of senescence-related secretory factors interleukin-1β (IL-1β) and interleukin-6 (IL-6) in the serum of mice. This composition is used for preventing or improving vascular aging, and has the characteristics of high efficiency, safety, and good stability. Brief Description of the Drawings

[0026] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for use in the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0027] Figure 1 For the P16 and P21 protein expression levels after Malabaricone C treatment of replicative senescent endothelial cells provided in Examples 1-4 of the present invention;

[0028] Figure 2 For the CCK-8 detection of the proliferation activity of senescent endothelial cells after Malabaricone C treatment of replicative senescence and pathological senescence provided in Examples 1-4 of the present invention;

[0029] Figure 3 For the accumulation level of β-galactosidase in senescent endothelial cells after Malabaricone C treatment of replicative senescence and pathological senescence provided in Example 1 of the present invention;

[0030] Figure 4 For the secretion levels of IL-1β and IL-6 in the serum after the action of the Malabaricone C complex on senescent mice provided in Examples 5-7 of the present invention; Figure 5The secretion level of NO in the serum after the Malabaricone C complex provided in Examples 5-7 of the present invention acts on senescent mice. Detailed implementation manners

[0031] Unless otherwise defined herein, scientific and technical terms used in conjunction with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. The meanings and scopes of the terms should be clear. However, in any case of potential ambiguity, the definitions provided herein shall prevail over any dictionary or extrinsic definition. In this application, unless otherwise specified, the use of "or" means "and / or". In addition, the use of the term "comprising" and other forms is non-limiting.

[0032] Generally, the nomenclatures and techniques used in conjunction with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those well known and commonly used in the art. Unless otherwise specified, the methods and techniques of the present invention are generally carried out according to conventional methods well known in the art and as described in various general and more specific references, which are cited and discussed throughout this specification. Enzymatic reactions and purification techniques are carried out according to the manufacturer's instructions, as commonly practiced in the art or as described herein. The nomenclatures used in conjunction with analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein, as well as the laboratory procedures and techniques, are those well known and commonly used in the art.

[0033] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts shall fall within the protection scope of the present invention.

[0034] The first aspect of the present invention provides an application of Malabaricone C in the preparation of a product for anti-vascular aging.

[0035] Furthermore, the effective substance concentration of Malabaricone C for improving the senescence of vascular endothelial cells is 0.5-20 μM.

[0036] More preferably, the effective concentration of Malabaricone C for improving the senescence of vascular endothelial cells is 10 μM.

[0037] Furthermore, Malabaricone C is used for improving the senescence of vascular endothelial cells, and the vascular endothelial cells include primary human umbilical vein endothelial cells (HUVEC).

[0038] In the second aspect of the present invention, an anti-vascular aging composition based on Malabaricone C is provided, and its components include Malabaricone C and optional excipients.

[0039] Specifically, the present invention provides a composition with Malabaricone C as the main component, containing PGG as the main active ingredient, as well as excipients acceptable in food, health products, and drugs.

[0040] Furthermore, the anti-vascular aging composition includes the following components by weight percentage:

[0041] 0.01% - 0.5% Malabaricone C and the balance of excipients.

[0042] Among them, "0.01% - 0.5%", for example, can be 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, etc. In the anti-vascular aging composition, the weight percentage of Malabaricone C is 0.01% to 0.5%.

[0043] Furthermore, the excipients may include excipients acceptable in food, health products, and drugs, and may also include other active ingredients, such as antioxidants, anti-inflammatory agents, etc.

[0044] Furthermore, the antioxidant is one or more of vitamin C and catechin; the anti-inflammatory agent is tea polyphenol.

[0045] In the third aspect of the present invention, an application of the anti-vascular aging composition in the preparation of food or health products is provided.

[0046] In the fourth aspect of the present invention, an application of the anti-vascular aging composition in the preparation of anti-vascular aging drugs is provided.

[0047] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0048] In the present invention, natural passage is used to simulate the replicative senescence of vascular endothelial cells, and glucose, hydrogen peroxide, and 5-fluorouracil are used to simulate the pathological senescence of vascular endothelial cells induced by high glucose, oxidative stress, and chemotherapy drugs respectively. Through the treatment with the compound Malabaricone C, it is found that it can significantly improve the natural senescence and pathological senescence of vascular endothelial cells. Specifically, the compound Malabaricone C significantly reduces the expression of cell cycle inhibitors P16 and P21 in vascular endothelial cells; at the same time, it also significantly reduces the presence of galactosidase staining deposits in endothelial cells.

[0049] The present invention uses 15-month-old wild-type C57B6 / J mice to establish a natural aging blood vessel model. During the experiment, by intraperitoneal injection of the compound Malabaricone C, it was found that it can improve the blood vessel aging induced by the natural aging model. The specific manifestations are as follows: The compound Malabaricone C can significantly reduce the expression levels of the senescence-related secretory factors interleukin-1β and interleukin-6 in the serum of mice. To further detect the effectiveness of the compound Malabaricone C in delaying blood vessel aging, the secretion level of NO in the serum of mice was analyzed, and it was found that the compound Malabaricone C can significantly enhance the secretion level of NO in the serum of mice.

[0050] The present invention first focuses on the anti-aging function of the compound Malabaricone C on the blood vessel system and verifies it using a reasonable and scientific experimental method.

[0051] The present invention will be further described below through examples. Unless otherwise specified, the materials in the examples are prepared according to existing methods or directly purchased from the market.

[0052] The CAS number of the compound Malabaricone C is 63335-25-1.

[0053] Example 1

[0054] The preparation process of the compound Malabaricone C working solution is as follows:

[0055] Accurately weigh 1.79215 mg of the compound Malabaricone C powder using an analytical balance and place it in a 1.5 mL sterile centrifuge tube. In a laminar flow hood, use a micropipette to add 1 mL of DMSO to the centrifuge tube, and repeatedly pipette with the pipette tip until the powder is completely dissolved. After the solution is clear without particles and impurities, a stock solution with a concentration of 5 mM is obtained. Then, it is aliquoted and stored in an ultra-low temperature refrigerator. When intervening with cells in vitro, take out the aliquoted compound Malabaricone C stock solution from the ultra-low temperature refrigerator, melt it in a 37°C water bath, then centrifuge at high speed, and place it in a laminar flow hood. The final concentration of the working solution for intervening with cells is 10 μM. The preparation process is as follows: According to the ratio of 1 mL of complete medium: 2 μL of Malabaricone C stock solution, use a micropipette to add the stock solution to the complete medium, mix well with the pipette tip and then add it to the cell culture dish. After the operation, place the culture dish in a cell culture incubator for incubation.

[0056] Example 2

[0057] In this example, the difference in the preparation process of the working solution of compound Malabaricone C from that of the example is as follows: the concentration of the working solution is different, and the final concentration of the working solution for intervening cells is 1 μM. The preparation process is as follows: according to the ratio of 1 mL of complete medium: 0.2 μL of Malabaricone C stock solution, use a micropipette to add the stock solution to the complete medium, and the remaining steps are the same as those in Example 1.

[0058] Example 3

[0059] The difference in the preparation of the working solution of compound Malabaricone C in Example 3 from that in Example 1 is as follows: the concentration of the working solution is different, and the final concentration of the working solution for intervening cells is 0.5 μM. The preparation process is as follows: according to the ratio of 1 mL of complete medium: 0.1 μL of Malabaricone C stock solution, use a micropipette to add the stock solution to the complete medium, and the remaining steps are the same as those in Example 1.

[0060] Example 4

[0061] The difference in the preparation of the working solution of compound Malabaricone C in Example 4 from that in Example 1 is as follows: the concentration of the working solution is different, and the final concentration of the working solution for intervening cells is 20 μM. The preparation process is as follows: according to the ratio of 1 mL of complete medium: 4 μL of Malabaricone C stock solution, use a micropipette to add the stock solution to the complete medium, and the remaining steps are the same as those in Example 1.

[0062] Example 5

[0063] A solvent containing Malabaricone C is prepared as follows:

[0064] Add 100 mg of Malabaricone C and 100 mg of cyclodextrin to about 8 mL of pure water, stir until completely dissolved; then add 10 mg of tea polyphenols and continue to stir until homogeneous; then add 50 mg of vitamin C and stir until completely dissolved; transfer the solution to a 10 mL volumetric flask, make up the volume to 10 mL with pure water, and shake gently. Finally, a homogeneous solution containing Malabaricone C (10 mg / mL), green tea extract (1 mg / mL), vitamin C (5 mg / mL), and cyclodextrin (10 mg / mL) is obtained.

[0065] Example 6

[0066] The difference in the preparation of the working solution of Malabaricone C in Example 6 from that in Example 5 is as follows: accurately weigh 50 mg of Malabaricone C, and the remaining steps are the same as those in Example 5. The final concentration of Malabaricone C in the composition is 5 mg / mL.

[0067] Example 7

[0068] The difference in the preparation of the working solution of Malabaricone C in Example 7 from that in Example 5 is as follows: Weigh 150 mg of Malabaricone C accurately, and the remaining steps are the same as those in Example 5. The concentration of Malabaricone C in the final composition is 15 mg / mL.

[0069] Comparative Example 1

[0070] The difference in the preparation of the working solution in Comparative Example 1 from that in Example 1 is as follows: Use dimethyl sulfoxide (DMSO) as the control working solution for Examples 1-4, and the volume used is the same as the volume of Malabaricone C used in Example 4.

[0071] Comparative Example 2

[0072] The difference in the preparation of the solvent in Comparative Example 2 from that in Example 5 is as follows: Use sterile normal saline as the control working solution for Examples 5-7, and the volume used is the same as the volume of the composition used in Example 7.

[0073] Test Examples 1-4: Detect the protein levels of P16 and P21 in senescent endothelial cells

[0074] Test Examples 1-4 were respectively tested using the working solutions prepared in Examples 1-4:

[0075] Primary human umbilical vein endothelial cells were passaged using ECM medium (37 °C, 5% CO2), and young endothelial cells were selected for the experiment (P3-P5). Through normal passaging, endothelial cells with P>15 were selected as replicative senescent cells. In the experiment, the cells were intervened with the compound Malabaricone C working solutions in Examples 1-4 respectively and maintained for 24 h. After the induction, the protein levels of P16 and P21 were detected by Western blot.

[0076] Control Test Example 1

[0077] The test was carried out using the working solution prepared in Comparative Example 1, and the test method was the same as that in Test Example 1.

[0078] The experimental results showed that: Referring to Figure 1 As can be seen, the compound Malabaricone C significantly reduced the protein levels of P16 and P21 in replicative senescent endothelial cells at 10 μM. P16 and P21 are considered as marker proteins of senescence. Therefore, the compound Malabaricone C can delay the senescence of endothelial cells.

[0079] Test Example 5 - 8: Effect of Compound Malabaricone C on the Proliferation of Senescent Endothelial Cells

[0080] Test Examples 5 - 8 were tested using the working solutions prepared in Examples 1 - 4 respectively:

[0081] Young endothelial cells were stimulated and induced by adding glucose (30 mM), H2O2 (200 μM), and 5 - fluorouracil (1 μM) to the culture medium respectively for 24 h to form senescent endothelial cells induced by high glucose, oxidative stress, and chemotherapeutic drugs respectively. Together with replicative senescent endothelial cells, subsequently, the working solutions of Compound Malabaricone C in Examples 1 - 4 were used to intervene in the cells for 48 h. Subsequently, the cell proliferation activity was detected by a CCK - 8 kit. Among them, in Test Example 5, the working solutions of Malabaricone C in Examples 1 - 4 were used to intervene in replicative senescent endothelial cells; in Test Example 6, the working solutions of Malabaricone C in Examples 1 - 4 were used to intervene in high - glucose senescent endothelial cells; in Test Example 7, the working solutions of Malabaricone C in Examples 1 - 4 were used to intervene in oxidative - stress senescent endothelial cells; in Test Example 8, the working solutions of Malabaricone C in Examples 1 - 4 were used to intervene in chemotherapeutic - drug - induced senescent endothelial cells.

[0082] Control Test Example 2

[0083] The test was carried out using the working solution prepared in Comparative Example 1, and the test method was the same as that in Test Example 5.

[0084] The experimental results are as Figure 2 shown: Compound Malabaricone C significantly enhanced the proliferation ability of replicative senescent( Figure 2 A) and oxidative - stress - damaged senescent( Figure 2 C) endothelial cells, while significantly inhibited the proliferation of high - glucose - induced( Figure 2 B) and chemotherapeutic - drug - damaged senescent endothelial cells( Figure 2 D). The experimental results suggest that the anti - endothelial - aging effect of 1,2,3,4,6 - pentagalloylglucose may exist in multiple ways of delaying aging or killing senescent cells.

[0085] Test Examples 9 - 12: Detection of Galactosidase Staining Deposits in Senescent Endothelial Cells

[0086] Test Examples 9 - 12 were tested using the working solution prepared in Example 1 respectively:

[0087] Young endothelial cells were stimulated and induced by adding glucose (30 mM), H2O2 (200 μM), and 5-fluorouracil (1 μM) to the culture medium respectively for 24 h to form senescent endothelial cells induced by high glucose, oxidative stress, and chemotherapeutic drugs. Together with replicative senescent endothelial cells, they were then intervened with the compound Malabaricone C working solution in Example 1 for 24 h. After the induction, they were stained with the galactosidase staining working solution and photographed. Scale bar, 200 μm. Among them, Test Example 9 used the Malabaricone C working solution in Example 1 to intervene in replicative senescent endothelial cells; Test Example 10 used the Malabaricone C working solution in Example 1 to intervene in high-glucose senescent endothelial cells; Test Example 11 used the Malabaricone C working solution in Example 1 to intervene in oxidative stress senescent endothelial cells; Test Example 12 used the Malabaricone C working solution in Example 1 to intervene in chemotherapeutic drug-induced senescent endothelial cells.

[0088] Control Test Example 3

[0089] The working solution prepared in Comparative Example 1 was used for testing, and the testing method was the same as that in Test Example 9.

[0090] The experimental results showed that: referring to Figure 3 it can be seen that the compound Malabaricone C significantly reduced the accumulation of galactosidase staining deposits in replicative senescent ( Figure 3 A and B) and high-glucose ( Figure 3 A and C), oxidative stress ( Figure 3 A and D), and chemotherapeutic drug-induced ( Figure 3 A and E) senescent endothelial cells. The galactosidase staining method is a currently recognized senescence detection method. According to the experimental results, the compound Malabaricone C can delay the replicative senescence and pathological senescence of endothelial cells, which is consistent with the experimental results at the Figure 1 molecular level.

[0091] Test Examples 13 - 15: Testing the secretion levels of IL-1β and IL-6 in mouse serum

[0092] Test Examples 13 - 15 were respectively tested using the solutions prepared in Examples 5 - 7:

[0093] The mice used in the experiment were all 15-month-old aged mice, with 5 mice in each group. In Test Examples 13-15, the Malabaricone C composite solution in Examples 5-7 was used to perform intraperitoneal injection on the mice once every 4 days. After continuous administration for four weeks, blood was taken from the orbital cavity, serum was separated, and the secretion levels of IL-1β and IL-6 in the mouse serum were detected.

[0094] Control Test Example 4

[0095] The solution prepared in Comparative Example 2 was used for testing, and the testing method was the same as that in Test Example 13.

[0096] The experimental results showed that referring to Figure 4 it could be known that the compound Malabaricone C significantly reduced the secretion levels of IL-1β ( Figure 4 A) and IL-6 ( Figure 4 B) in the mouse serum. Senescent cells can secrete a variety of factors to further promote the senescence of the body, and senescence-associated secretory factors are important markers for evaluating the senescence of the body. After the intervention of the compound Malabaricone C, the secretion levels of IL-1β and IL-6 in the mouse serum could be significantly reduced. Combining the above experimental results, it could be known that the compound Malabaricone C could also delay senescence in mice in vivo, which was consistent with the anti-aging effect of in vitro cell experiments.

[0097] Test Examples 16-18: Testing the NO level in the serum of senescent mice

[0098] Test Examples 16-18 were respectively tested with the solutions prepared in Examples 5-7:

[0099] The mice used in the experiment were all 15-month-old aged mice, with 5 mice in each group. In Test Examples 16-18, the Malabaricone C composite solution in Examples 5-7 was used to perform intraperitoneal injection on the mice once every 4 days. After continuous administration for four weeks, blood was taken from the orbital cavity, serum was separated, and the secretion level of NO in the mouse serum was detected.

[0100] Control Test Example 5

[0101] The solution prepared in Comparative Example 2 was used for testing, and the testing method was the same as that in Test Example 16.

[0102] The experimental results showed that referring to Figure 5 it could be known that compared with the control group, the compound Malabaricone C significantly increased the NO level in the serum of senescent mice after treatment. This indicated that the compound Malabaricone C could effectively improve the vascular function in vivo.

[0103] The experiments and results of the present invention have proved that the compound Malabaricone C can effectively inhibit the senescence of vascular endothelial cells and vascular aging.

[0104] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of Malabaricone C in the preparation of a product for anti-vascular aging.

2. The application according to claim 1, wherein The effective substance concentration of Malabaricone C for improving the senescence of vascular endothelial cells is 0.5 - 20 μM.

3. The application according to claim 1, wherein The effective concentration of Malabaricone C for improving the senescence of vascular endothelial cells is 10 μM.

4. The application according to claim 1, characterized in that Malabaricone C is used for improving the senescence of vascular endothelial cells, and the vascular endothelial cells include primary human umbilical vein endothelial cells.

5. An anti-vascular aging composition based on Malabaricone C, characterized in that, Its components include Malabaricone C and optional excipients.

6. The anti-aging blood vessel composition according to claim 5, characterized in that, The anti-vascular aging composition comprises the following components by weight percentage: 0.01% - 0.5% Malabaricone C and the balance of excipients.

7. The anti-aging vascular composition according to claim 5, wherein The excipients include one or more of antioxidants and anti-inflammatory agents.

8. The anti-aging composition according to claim 7, wherein The antioxidants include one or more of vitamin C and catechins; The anti-inflammatory agent includes tea polyphenols.

9. Use of the anti-vascular aging composition according to any one of claims 5 - 8 in the preparation of a food or health product.

10. Use of the anti-vascular aging composition according to any one of claims 5 - 8 in the preparation of an anti-vascular aging drug.

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