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Title:
ANTI-AGING METHODS AND COMPOSITIONS
Document Type and Number:
WIPO Patent Application WO/2019/104065
Kind Code:
A1
Abstract:
The present application provides methods, compositions, and kits for treating or preventing an aging-related disease or condition in an individual. The method comprises administration of a combination regimen, including an insulin sensitizer, an mTOR inhibitor, or an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; a senolytic p53/pRb activator; and an NAD+ supplement. Each component of the combination regimen may be administered systemically or locally. Many clinically available pharmaceuticals are suitable for use in this combination regimen.

Inventors:
YEUNG, Alex Wah Hin (8093 Scholarship, Irvine, California, 92612, US)
Application Number:
US2018/062067
Publication Date:
May 31, 2019
Filing Date:
November 20, 2018
Export Citation:
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Assignee:
TURRINII PHARMACEUTICAL, LLC (8093 Scholarship, Irvine, California, 92612, US)
International Classes:
A61K31/195; A61P25/28
Domestic Patent References:
WO2017062311A12017-04-13
Foreign References:
US20120283269A12012-11-08
US20020128278A12002-09-12
US20030170287A12003-09-11
US5536729A1996-07-16
US20060229265A12006-10-12
Other References:
LEONTIEVA ET AL.: "Gerosuppression by pan-mTOR inhibitors", AGING, vol. 8, no. 12, 30 December 2016 (2016-12-30), pages 3535 - 3549, XP055615511
PANTZIARKA ET AL.: "Repurposing Drugs in Oncology (ReDO) - diclofenac as an anti-cancer agent", ECANCERMEDICALSCIENCE, vol. 10, 2016, pages 1 - 32, XP002791052
PALMIERI ET AL.: "Ultrasound Lipolysis and Suction Lipectomy for Treatment of Obesity", THE AMERICAN JOURNAL OF COSMETIC SURGERY, vol. 14, no. 3, 1 September 1997 (1997-09-01), pages 289 - 296, XP055615514
CHOI ET AL.: "Cyclooxygenase-2 functionally inactivates p53 through a physical interaction with p53", BIOCHIMICA ET BIOPHYSICA ACTA (BBA) - MOLECULAR CELL RESEARCH, vol. 1793, no. 8, 2009, pages 1354 - 1365, XP026336410, doi:10.1016/j.bbamcr.2009.05.006
DING ET AL.: "Current Perspective in the Discovery of Anti-aging Agents from Natural Products", NATURAL PRODUCTS AND BIOPROSPECTING, vol. 7, no. 5, 31 May 2017 (2017-05-31) - October 2017 (2017-10-01), pages 335 - 404, XP055615517
Attorney, Agent or Firm:
QI, Yan et al. (Morrison & Foerster LLP, 755 Page Mill RoadPalo Alto, California, 94304-1018, US)
Download PDF:
Claims:
CLAIMS

What is claimed is:

1. A method of treating or preventing an aging-related disease or condition in an individual, comprising administering to the individual:

(i) an effective amount of an insulin sensitizer and an effective amount of an mTOR inhibitor, or an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) an effective amount of a senolytic p53/pRb activator; and

(iii) an effective amount of an NAD+ supplement.

2. The method of claim 1, comprising administering to the individual an effective amount of an insulin sensitizer and an effective amount of an mTOR inhibitor.

3. The method of claim 2, wherein the insulin sensitizer comprises a thiazolidinedione

and/or a biguanide.

4. The method of claim 3, wherein the insulin sensitizer comprises a thiazolidinedione, wherein the thiazolidinedione is pioglitazone.

5. The method of claim 3, wherein the insulin sensitizer comprises a biguanide, wherein the biguanide is metformin.

6. The method of claim 2, wherein the mTOR inhibitor comprises rapamycin and/or a

rapalog.

7. The method of claim 2, wherein the mTOR inhibitor comprises a pan-mTOR inhibitor.

8. The method of claim 1, comprising administering to the individual an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor.

9. The method of claim 8, wherein the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin.

10. The method of claim 1, wherein the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor, a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof.

11. The method of claim 10, wherein the senolytic p53/pRb activator comprises a COX

inhibitor, and wherein the COX inhibitor is diclofenac.

12. The method of claim 1, wherein the NAD+ supplement comprises Nicotinic Acid (NA), Nicotinamide Riboside (NR), Nicotinamide Mononucleotide (NMN), and/or

Nicotinamide (NAM).

13. The method of claim 12, wherein the NAD+ supplement comprises NR.

14. The method of claim 1, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are nutraceutical agents.

15. The method of claim 1, wherein the aging-related disease or condition is selected from the group consisting of cancer, cardiovascular diseases, central nervous system diseases, Alzheimer's disease, Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, osteoporosis, arthritis, insulin-resistance and type II diabetes, benign prostatic hyperplasia, systemic lupus erythematosus, auto-immune diseases, psoriasis, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer, eczema, pain, inflammation or strain of muscle, tendon or joint, fat management related to lipectomy, liposuction, lipolysis and fat sculpturing from cold, heat, laser, ultrasound and radiofrequency ablation therapy, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, arthritis, and combinations thereof.

16. The method of claim 1, wherein the method is for cosmetic treatment.

17. The method of claim 1, wherein the aging-related disease or condition is obesity, non alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, pre-diabetic, diabetes type II and other fat or inflammatory induced metabolic imbalances.

18. The method of claim 1, wherein the individual further receives a fat reduction by

lipectomy or modification by a lipolysis procedure.

19. The method of claim 1, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered systemically.

20. The method of claim 1, wherein the insulin sensitizer, mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement is administered via a combination of local and systemic routes.

21. The method of claim 1, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered locally.

22. The method of claim 21, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered via an implanted device.

23. The method of claim 22, wherein the implanted device is selected from the group

consisting of an intravesical or other urogenital tract gel, an intra-cranial or intra-thecal device, intra-ocular device, a drug-eluting stent, and CT, MRI or X-ray guided and inserted devices.

24. The method of claim 1, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is formulated in a pharmaceutical composition comprising a pharmaceutically acceptable excipient.

25. The method of claim 24, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and the NAD+ supplement are formulated in a single pharmaceutical composition.

26. The method of claim 24, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions.

27. The method of claim 25, wherein the pharmaceutically acceptable excipient is a serum, gel, buffer solution, cream, lotion, liniment, ointment or combinations thereof.

28. The method of claim 25, wherein the agent having dual functions as an insulin sensitizer and an mTOR inhibitor is in a pharmaceutical composition comprising at least about 0.5% (w/v) metformin.

29. The method of claim 25, wherein the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0.5% (w/v) diclofenac.

30. The method of claim 25, wherein the NAD+ supplement is in a pharmaceutical

composition comprising at least 0.5 % (w/v) nicotinamide riboside.

31. The method of claim 25, wherein the NAD+ supplement is in a pharmaceutical

composition comprising at least 0.5 % (w/v) nicotinic acid.

32. The method of claim 25, wherein the pharmaceutical composition comprises about 0.5- 10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) nicotinamide riboside, about 0.5-6% (w/v) nicotinic acid, and about 0.5-10% (w/v) pterostilbene.

33. The method of claim 32, wherein the pharmaceutical composition further comprises about 1-10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40%ϊ (w/v) PEG200, and about 5-40% (w/v) PEG400.

34. The method of claim 21, wherein the pharmaceutical composition is administered to the individual topically.

35. The method of claim 21, wherein the aging-related disease or condition is an allergic or auto-immune skin disease, and wherein the method further comprises locally

administering an effective amount of a corticosteroid to the individual.

36. The method of claim 1, wherein the method further comprises administering to the

individual an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-camitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Cbayote (, Seckhim edide) juice or extract.

37. The method of claim 1, wherein the individual is a human.

38. The method of claim 1, wherein the individual is a pet.

39. A method of decelerating, stabilizing, ameliorating, or reversing one or more signs of aging in a mammalian cell, comprising contacting the mammalian cell with:

(i) an effective amount of an insulin sensitizer and an effective amount of an mTOR inhibitor, or an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) an effective amount of a senolytic p53/pRb activator; and

(iii) an effective amount of an NAD+ supplement.

40. A pharmaceutical composition comprising:

(i) an insulin sensitizer and an mTOR inhibitor, or an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) a senolytic p53/pRb activator; and

(iii) an NAD+ supplement;

and a pharmaceutically acceptable excipient.

41. A pharmaceutical composition comprising about 0.5-10% (w/v) metformin, about 0.5- 4% (w/v) diclofenac, about 0.5-6% (w/v) nicotinamide riboside, about 0.5-6% (w/v) nicotinic acid, about 0.5-10% (w/v) pterostilbene, and a pharmaceutically acceptable excipient.

42. The pharmaceutical composition of claim 41, wherein the pharmaceutical composition is formulated for topical administration.

43. The pharmaceutical composition of claim 41, further comprising about 1-10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40% (w/v) PEG200, and about 5-40% (w/v) PEG400.

44. A kit for treating or preventing an aging-related disease or condition, comprising:

(i) an insulin sensitizer and an mTOR inhibitor, or an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) a senolytic p53/pRb activator; and

(iii) an NAD+ supplement.

45. An implantable device for treating or preventing an aging-related disease or condition, comprising:

(i) an insulin sensitizer and an mTOR inhibitor, or an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) a senolytic p53/pRb activator, and

(iii) an NAD+ supplement.

Description:
ANTI-AGING METHODS AND COMPOSITIONS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 62/590,138 filed on November 22, 2017, the contents of which are incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to methods, compositions and kits for treating or preventing aging-related diseases or conditions.

BACKGROUND OF THE INVENTION

[0003] Aging in humans is a well-established risk factor for many disabling diseases and conditions, including diabetes, cardiovascular diseases, Alzheimer’s disease and cancer. In fact, the risk of death from these causes is dramatically accelerated by 100-1000 fold between the ages of 35 and 85 years. For this reason, there is a need for the development of new

interventions to improve and maintain health into old age, i.e., to improve“health span” in order to obtain a healthy end result for aging

[0004] Several mechanisms have been shown to delay the aging process, resulting in improved lifespan in animal models. These mechanisms include caloric restriction, alteration in GH/IGF1 pathways, activation of sirtuins, and mTOR inhibition. Monotherapy agents, such as resveratrol (SIRTI activator) and rapamycin or a rapamycin-like nutraceutical agent (mTOR inhibitor), have been tested for anti -aging applications.

[0005] The disclosures of ail publications, patents, patent applications and published patent applications referred to herein are hereby incorporated herein by reference in their entirety.

BRIEF SUMMARY OF THE INVENTION

[0006] The present application provides methods, compositions, and kits comprising an insulin sensitizer, an mTOR inhibitor, or an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; a senolytic p53/pRb activator, and an NAD+ supplement for treating or preventing an aging-related disease or condition in an individual. Some embodiments of the methods combine pharmaceutical agents that are already in common clinical use, including FDA approved drugs, to achieve synergistic anti-aging results. The methods described herein can stabilize, suppress, ameliorate or even reverse signs of aging.“Aging” as used herein refers to organismal aging and/or cellular aging, including diseases or symptoms arising from aging.

[0007] One aspect of the present application provides a method of treating or preventing an aging-related disease or condition in an individual (e.g, human or pet), comprising

administering to the individual: (i) an effective amount of an insulin sensitizer; (ii) an effective amount of an mTOR inhibitor; (iii) an effective amount of a senolytic p53/pRb activator; and (iv) an effective amount of an NAD+ supplement. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered systemically. In some embodiments, the insulin sensitizer, mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement is administered via a combination of local and systemic routes. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered locally.

[0008] In some embodiments according to any one of the methods described above, the aging- related disease or condition is selected from the group consisting of cancer (e.g., for prevention or as adjuvant therapy), cardiovascular diseases, central nervous system diseases, Alzheimer's disease, Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, osteoporosis, arthritis, insulin-resistance and type II diabetes, benign prostatic hyperplasia, systemic lupus erythematosus, auto-imnume diseases, psoriasis, and combinations thereof. In some embodiments, the aging-related disease or condition is selected from the group consisting of obesity, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, pre- diabetic, type II diabetes and other fat or inflammatory induced metabolic imbalances (e.g · ., hypercholesterolemia).

[0009] In some embodiments according to any one of the methods described above, the method is for cosmetic treatment. In some embodiments, the individual further receives a fat reduction by lipectomy or modification by a lipolysis procedure (such as the surgical procedure of liposuction and/or application of cold, heat, laser, ultrasound, radiofrequency ablation for lipolysis and fat sculpturing, e.g., using machines used such as but not exclusive limited to,

Cool Sculpture, Cynosure SculpSure and similar lipolysis machines from other manufacturers).

[0010] In some embodiments according to any one of the methods described above, wherein the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered locally, the aging-related disease or condition is selected from the group consisting of primary or metastatic cancer lesions (e.g, for prevention or as adjuvant therapy), cardiovascular diseases, central nervous system diseases, Alzheimer's disease,

Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g, for prevention or as adjuvant therapy), psoriasis, eczema, pain, inflammation or strain of muscle, tendon or joint, lipolysis (e.g, by cold, heat, laser, ultrasound, or radiofrequency ablation for fat reduction and modification as a cosmetic procedure or a treatment for obesity), post lipectomy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, and combinations thereof. In some embodiments, the insulin sensitizer, the mTGR inhibitor, the senolytic p53/pRb activator, and/or the NL1) · supplement is administered via an implanted device. In some embodiments, the implanted device is selected from the group consisting of an intravesical or other urogenital tract gel, an intra-cranial or intra-thecal device, intra-ocular device, a drug-eluting stent, and CT, MRI or X-ray guided and inserted devices. In some embodiments, the aging-related disease or condition is an allergic or auto-immune skin disease, and wherein the method further comprises locally administering an effective amount of a corticosteroid to the individual.

[0011] In some embodiments according to any one of the methods described above, the method further comprises administering to the individual an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-carnitine, R-lipoic acid, leptine, a caspase inhibitor, and/or an antioxidant compound (e.g., Chayote (Sechium edii!e) juice or extract).

[0012] Another aspect of the present application provides a method of decelerating, stabilizing, ameliorating, or reversing one or more signs of aging in a mammalian cell, comprising contacting the mammalian cell with: (i) an effective amount of an insulin sensitizer; (ii) an effective amount of an mTOR inhibitor; (iii) an effective amount of a senolytic p53/pRb activator; and (iv) an effective amount of an NAD+ supplement.

[0013] In some embodiments according to any one of the methods described above, the method comprises administering an agent having dual functions selected from the group consisting of insulin sensitizer, mTOR inhibitor, senolytic p53/pRb activator, and NAIH supplement. In some embodiments, the method comprises administering to the individual: (i) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (ii) an effective amount of a senolytic p53/pRb activator; and (iii) an effective amount of an NAD+ supplement. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor is metformin. In some embodiments, the agent is administered systemicaily. In some embodiments, the agent is administered locally.

[0014] In some embodiments according to any one of the methods described above, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are nutraceutical agents.

[0015] In some embodiments according to any one of the methods described above, the insulin sensitizer comprises a thiazolidinedione and/or a biguanide. In some embodiments, the thiazolidinedione is pioglitazone. In some embodiments, the biguanide is metformin. In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent.

[0016] In some embodiments according to any one of the methods described above, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog, such as everolimus, tacrolimus, CCI-779, ABT-578, AP-23675, AP-23573, AP-23841, 7-epi -rapamycin, 7-thiomethyl -rapamycin, 7-epi -trimethoxyphenyl -rapamycin, 7-epi-thiomethyl-rapamycin, 7- demethoxy-rapamycin, 32-demethox -rapamycin, 2-rapamycin, 7-demethoxy-rapamycin, 32- demethoxy-rapamycin, 2-desmethyl-rapamycin, 42-0-(2-hydroxy)ethyl rapamycin, or any combination thereof. In some embodiments, the effective amount of rapamycin or the rapalog maintains a trough blood level of rapamycin selected from no more than about 8ng/m! for systemic administration, no more than about 2ng/m! for local administration, and no more than about lOng/mi for combined systemic and local administration, or an equivalent trough blood level of the rapalog thereof In some embodiments, the mTOR inhibitor comprises a pan-mTQR inhibitor, such as sapanisertib (INK128), AZD2014, torin 1, torin 2, AZD8055, PP242, KU- 006379, 081-027, WAY-600, WAE-687, WYE-354, GSK1059615, or any combination thereof [0017] In some embodiments according to any one of the methods described above, the senolytic p53/pRb activator is selected from a nutlin or analog thereof a COX inhibitor, a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, wherein the senolytic p53/pRb activator comprises a nutlin, the nutlin is nutlin-3A. In some embodiments, wherein the senolytic p53/pRb activator comprises a COX inhibitor, the COX inhibitor is selected from the group consisting of aspirin, ibuprofen, naproxen, diclofenac, indomethacin, and combinations thereof In some embodiments, the senolytic p53/pRb activator is diclofenac. In some embodiments, wherein the senolytic p53/pRb activator comprises a pan CDK inhibitor, the pan CDK inhibitor is selected from the group consisting of flavopiridol, o!omoucine II, purvalanol A, SNS-032, dinaciclib, MK-7965, SCH727965, AT7519, R547, AZD5438, and AG024322. In some embodiments, wherein the senolytic p53/pRb activator comprises a CDK4/6, the CDK4/6 inhibitor is selected from the group consisting of ribocicfib, palbociclib, abemaciclib, and trilaciclib. In some embodiments, the senolytic p53/pRb activator comprises any combination of p53 activators, COX inhibitors, pan CDK inhibitors, and CDK4/6 inhibitors.

[0018] In some embodiments according to any one of the methods described above, the NAD+ supplement comprises Nicotinic Acid (NA), Nicotinamide Riboside (NR), Nicotinamide Mononucleotide (NMN), and/or Nicotinamide (NAM). In some embodiments, the NAD+ supplement comprises: (a) NA and (b) one or more of NR, NMN and NAM. In some embodiments, the NAD+ supplement comprises NR and NA. In some embodiments, the effective amount of the N AD supplement maintains a blood level of at least about 500mM NAD -

[0019] In some embodiments according to any one of the methods described above, the insulin sensitizer, the mTQR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is formulated in a pharmaceutical composition comprising a pharmaceutically acceptable excipient. In some embodiments, the insulin sensitizer and the mTOR inhibitor, or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor; the senolytic p53/pRb activator; and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an ni ' TOR inhibitor, the senolytic p53/pRb activator, and the NAD+ supplement are each formulated in a different matrix for immediate or extended release. In some embodiments, the pharmaceutically acceptable excipient is a serum, gel, buffer solution, cream, lotion, liniment, ointment or combinations thereof. In some embodiments, the insulin sensitizer or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor is in a pharmaceutical composition comprising at least about 0.5% (w/v; e.g., at least about 0.7%) metformin or an insulin sensitizing nutraceutical agent. In some embodiments, the insulin sensitizer is in a pharmaceutical composition comprising at least about 0.03% (w/v) pioglitazone. In some embodiments, the mTOR inhibitor is in a pharmaceutical composition comprising at least about 0.01% (w/v) rapamycin or a rapamycin-like nutraceutical agent. In some embodiments, the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 1% (w/v) aspirin. In some embodiments, the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0.5% (w/v) diclofenac. In some embodiments, the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0.3% (w/v) nutlin-3. In some embodiments, the NAD+ supplement is in a pharmaceutical composition comprising: (a) at least about 0.5% (w/v) Nicotinic Acid, and (b) at least about 1% (w/v)

Nicotinamide Riboside (NR), at least about 1.5% (w/v) Nicotinamide Mononucleotide ( MN), and/or at least about 0.5% (w/v) Nicotinamide (NAM). In some embodiments, wherein the NAD+ supplement is in a pharmaceutical composition comprising at least 0.5 % (w/v) NR.

[0020] In some embodiments according to any one of the methods described above, the method comprises administering to the individual an effective amount of a pharmaceutical composition comprising: about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) nicotinamide riboside, about 0.5-6% (w/v) nicotinic acid, about 0.5-10% (w/v) pterosti!bene, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises about 1 -10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40% (w/v) PEG200, and about 5-40% (w/v) PEG400. In some embodiments, the pharmaceutical composition is administered to the individual topically. In some embodiments, the pharmaceutical composition comprises about 2% (w/v) diclofenac, about 2% (w/v) metformin, about 3% (w/v) NR and about 1.5% (w/v) pterostilbene. In some embodiments, the pharmaceutical composition further comprises about 0 5-6% (w/v) NA, such as about 1.5% (w/v) NA.

[0021] One aspect of the present application provides a pharmaceutical composition

comprising: (i) an insulin sensitizer (e.g. , metformin or an insulin sensitizing nutraceutical agent); (ii) an mTOR inhibitor (e.g., rapamycin or a rapamycin-like nutraceutical agent, or torin 2), (iii) a senolytic p53/pRb activator (e.g., ibuprofen); (iv) an NAD+ supplement (e.g, NA and NR); and a pharmaceutically acceptable excipient.

[0022] In some embodiments, there is provided a pharmaceutical composition comprising: (i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor (e.g., metformin), (ii) a senolytic p53/pRb activator (e.g., diclofenac); (iii) an NAD+ supplement (e.g., NR); and a pharmaceutically acceptable excipient. [0023] In some embodiments, there is provided a pharmaceutical composition comprising: about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) NR, about 0.5-6% (w/v) NA, about 0.5-10% (w/v) pterostilbene, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises about 1-10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1 -30% (w/v) ethanol, about 5-40% (w/v) PEG200, and about 5-40% (w/v) PEG400. In some embodiments, the pharmaceutical

composition is formulated for topical administration. In some embodiments, the pharmaceutical composition comprises about 2% (w/v) diclofenac, about 2% (w/v) metformin, about 3% (w/v)

NR and about 1.5% (w/v) pterostilbene. In some embodiments, the pharmaceutical composition further comprises about 0.5-6% (w/v) NA, such as about 1.5% (w/v) NA.

[0024] Also provided is a kit for treating or preventing an aging-related disease or condition, comprising: (i) an insulin sensitizer (e.g., metformin or an insulin sensitizing nutraceutical agent); (ii) an mTOR inhibitor (e.g, rapamycin or a rapamycin-like nutraceutical agent, or torin 2); (iii) a senolytic p53/pRb activator (e.g, ibuprofen); and (iv) an NAD+ supplement (e.g., NA and NR). In some embodiments, there is provided a kit comprising (i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor (e.g., metformin), (ii) a senolytic p53/pRb activator (e.g, diclofenac), and (iii) an NAD+ supplement (e.g, NR).

[0025] Further provided is an implantable device for treating or preventing an aging-related disease or condition, comprising: (i) an insulin sensitizer (e.g., metformin or an insulin sensitizing nutraceutical agent): (ii) an mTOR inhibitor (e.g., rapamycin or a rapamycin-like nutraceutical agent, or torin 2); (iii) a senolytic p53/pRb activator (e.g, ibuprofen); and (iv) an NAD+ supplement (e.g, NA and NR). In some embodiments, there is provided an implantable device comprising: (i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor (e.g, metformin), (ii) a senolytic p53/pRb activator (e.g, diclofenac), and (iii) an NAD+ supplement (e.g, NR).

[0026] These and other aspects and advantages of the present invention will become apparent from the subsequent detailed description and the appended claims. It is to be understood that one, some, or all of the properties of the various embodiments described herein may be combined to form other embodiments of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIG. 1 shows results of Patient 1 on the systemic Trial A.

[0028] FIG. 2 shows results of Patient 2 on the systemic Trial A. [0029] FIG. 3 shows results of Patient 1 on the local Trial C.

DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention provides methods, compositions and kits for treating or preventing an aging-related disease or condition in an individual, comprising a combination regimen, including an insulin sensitizer and an mTQR inhibitor, or an agent having dual functions as an insulin sensitizer and an mTQR inhibitor; a seno!ylic p53/pRb activator; and an NAD+ supplement. The safety and efficacy of the combination regimen is supported by data from the clinical trial described herein.

[0031] Recent research for anti-aging interventions is usually directed towards one particular mode of action. However, aging involves hundreds to thousands of molecules that have different effects on many aspects of bodily functions and their respective metabolic pathways. After years of slow degradation due to numerous perturbations to these molecular players, a progression to senescence and death becomes inevitable. Without being bound by any theory or hypothesis, the methods described herein, also referred as“combination regimen,” can lower the rate of aging and ameliorate its ill results. Any two of the four components (insulin sensitizer, mTQR inhibitor, senolytic p53/pRb activator, and NAD+ supplement) can comprise of, or being represented by, only one agent with dual component properties, thereby reducing the 4- component formulation into a 3-component formulation. Additionally, any one or more, or all of the four components can comprise of, or being represented by nutraceutical agents with the same component properties. In particular, the insulin sensitizer can maintain or revive insulin sensitivity, which can lead to a metabolic shift critical to basic cellular survival, functions and preservation. The mTQR inhibitor is designed to target one of the most pivotal axes for aging: mTQR is activated by default throughout one’s lifespan, while its activity is accentuated during active growth and feeding. The senolytic p53 inhibitor and the NAD+ supplement can promote DMA repair as well as improve the senescent and inflammatory environment from chronic

TQR activation. During aging, the senescent and inflammatory environment may be established in the body systemically, or localized in a specific tissue or organ, which may cause age-related diseases or conditions. The combination regimen described herein is applicable for both systemic and/or local treatment. Notably, the present invention can be practiced using some of the clinically available pharmaceuticals currently on the market.

[0032] Accordingly, one aspect of the present application provides a method of treating or preventing an aging-related disease or condition in an individual, comprising administering to the individual : (i) an effective amount of an insulin sensitizer; (ii) an effective amount of an mTGR inhibitor; (iii) an effective amount of a senolytic p53/pRb activator; and (iv) an effective amount of an NAD+ supplement. In some embodiments, the present application provides a method of treating or preventing an aging-related disease or condition in an individual, comprising administering to the individual : (i) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor (e.g., metformin); (ii) an effective amount of a senolytic p53/pRb activator (e.g., diclofenac), and (iii) an effective amount of an NAD+ supplement (e.g, NR).

[0033] Also provided are compositions, kits and articles manufacture useful for the methods described herein.

I. Definitions

[0034] Unless otherwise defined, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context or expressly indicated, singular terms shall include pluralities and plural terms shall include the singular.

[0035] The term“combination regimen” is used herein to refer to anti-aging therapies described herein comprising: an insulin sensitizer, an mTOR inhibitor, a senolytic p53/pRb activator, and a NAD+ supplement. Each of components in this combination regimen may comprise a single agent, or two or more agents. Any two of the components in this combination regimen can comprise of, or being represented by, only one agent with dual component properties, thereby reducing the 4-component formulation into a 3-component formulation. Any one or more, or all of the components in this combination regimen can comprise of, or being represented by nutraceutical agents with the same component properties

[0036] As used herein,“treatment” or“treating” is an approach for obtaining beneficial or desired results including clinical results. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms resulting from an aging-related disease or condition, diminishing the extent of the aging-related disease or condition, stabilizing the aging-related disease or condition (e.g, preventing or delaying the aging-related disease or condition), preventing or delaying the spread of the aging-related disease or condition, preventing or delaying the recurrence of the aging- related disease or condition, reducing recurrence rate of the aging-related disease or condition, delay or slowing the progression of the aging-related disease or condition, ameliorating the aging-related disease or condition, providing a remission (partial or total) of the aging-related disease or condition, decreasing the dose of one or more other medications required to treat the aging-related disease or condition, delaying the progression of the aging-related disease or condition, increasing the quality of life, and/or prolonging survival. In some embodiments, the treatment reduces the severity of one or more symptoms associated with aging-related disease or condition by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 100% compared to the corresponding symptom in the same subject prior to treatment or compared to the corresponding symptom in other subjects not receiving the treatment. Also encompassed by "treatment" is a reduction of pathological consequence of an aging-related disease or condition. The methods of the invention contemplate any one or more of these aspects of treatment.

[0037] As used herein,“delaying” the development of an aging-related disease or condition means to defer, hinder, slow, retard, stabilize, and/or postpone development of the disease or condition. This delay can be of varying lengths of time, depending on the history of the disease and/or individual being treated. As is evident to one skilled in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease. A method that“delays” development of an aging-related disease or condition is a method that reduces probability of disease development in a given time frame and/or reduces the extent of the disease in a given time frame, when compared to not using the method. Such comparisons are typically based on clinical studies, using a statistically significant number of subjects.

[0038] “Preventing,” as used herein, includes providing prophylaxis with respect to the occurrence or recurrence of an aging-related disease or condition in a subject that may be predisposed to the disease or condition but has not yet been diagnosed with the disease or condition.

[0039] An“effective amount” of an agent refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result, such as ameliorate, palliate, lessen, and/or delay one or more of its symptoms. The effective amount of an agent depends on the route of administration (e.g., systemic and/or local administration) of the agent. For some agents, an effective agent is chosen by monitoring the trough level, AUC level and/or a pharmacokinetically defined level, e.g., serum or urine level, of the agent and/or its metabolites. [0040] A“therapeutically effective amount” of an agent may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the agent to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the agent are outweighed by the therapeutically beneficial effects. A therapeutically effective amount may be delivered in one or more administrations.

[0041] A“prophyiacticaliy effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically but not necessarily, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophyiacticaliy effective amount will be less than the therapeutically effective amount.

[0042] As is understood in the art, an“effective amount” may be in one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired treatment endpoint. An effective amount may be considered in the context of administering one or more therapeutic agents. For example, any component of the combination regimen described herein may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable or beneficial result may be or is achieved. The components in a combination regimen of the invention may be administered sequentially, simultaneously, or continuously using the same or different routes of administration and the same or different schedule for each component. Thus, an effective amount of a combination regimen includes an amount of the insulin sensitizer, an amount of the mTOR inhibitor, or an amount of the agent having dual functions as an insulin sensitizer and an mTOR inhibitor: an amount of the senolytic p53/pRb activator, and an amount of the NAD supplement when administered sequentially, simultaneously, or continuously produces a desired outcome.

[0043] Administration“in combination with” one or more further therapeutic agents includes simultaneous (concurrent) and consecutive or sequential administration in any order.

[0044] The term“concurrently” and“simultaneously” are used herein to refer to

administration of two or more therapeutic agents, where at least part of the administration overlaps in time or where the administration of one therapeutic agent falls within a short period of time relative to administration of the other therapeutic agent. For example, the two or more therapeutic agents are administered with a time separation of no more than about 60 minutes, such as no more than about any of 30, 15, 10, 5, or 1 minute.

[0045] The term“sequentially” is used herein to refer to administration of two or more therapeutic agents where the administration of one or more agent(s) continues after discontinuing the administration of one or more other agent(s). For example, administration of the two or more therapeutic agents are administered with a time separation of more than about 15 minutes, such as about any of 20, 30, 40, 50, or 60 minutes, 1 day, 2 days, 3 days, 1 week, 2 weeks, or 1 month, or longer.

[0046] The term“individual,”“subject,” and“patient” are used interchangeably herein to describe a mammal, including humans. An individual includes, but is not limited to, human, bovine, horse, feline, canine, rodent, or primate. In some embodiments, the individual is human. In some embodiments, the individual is in need of treatment. In some embodiments, the individual is a sport animal, or a pet animal.

[0047] The terms“pharmaceutical formulation” and“pharmaceutical composition” refer to a preparation which is in such form as to permit the biological activity of the active ingredient(s) to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. Such formulations may be sterile.

[0048] A“pharmaceutically acceptable carrier” refers to a non-toxic solid, semisolid, or liquid filler, diluent, encapsulating material, formulation auxiliary, or carrier conventional in the art for use with a therapeutic agent that together comprise a“pharmaceutical composition” for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation. The pharmaceutically acceptable carrier is appropriate for the formulation employed.

[0049] It is understood that embodiments of the invention described herein include“consisting” and/or“consisting essentially of” embodiments.

[0050] Reference to "about" a value or parameter herein includes (and describes) variations that are directed to that value or parameter per se. For example, description referring to "about X" includes description of "X".

[0051] As used herein, reference to "not" a value or parameter generally means and describes "other than" a value or parameter.

[0052] The term“about X-Y” used herein has the same meaning as“about X to about Y.”

[0053] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. II. Combination regimen

[0054] The present application provides an anti-aging combination therapy comprising: an insulin sensitizer, an mTOR inhibitor, a senolytic p53/pRb activator, and a NAD+ supplement.

In some embodiments, an agent having dual functions is used. For example, in some

embodiments, the anti-aging combination therapy comprises an agent having dual functions as an insulin sensitizer and an mTOR inhibitor, a senolytic p53/pRb activator, and an NAD+ supplement. Methods, compositions, and kits of the combination therapy are provided. The combination therapy is also referred herein as“combination regimen.”

[0055] In some embodiments, there is provided a method of treating or preventing an aging- related disease or condition in an individual (e.g, a human or a pet animal), comprising administering to the individual: (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ {e.g., piog!itazone) and/or a biguanide {e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog {e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan-mTQR inhibitor {e.g., form 2). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g, ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the aging-related disease or condition is selected from the group consisting of cancer (e.g., for prevention or as adjuvant therapy), cardiovascular diseases, central nervous system diseases, Alzheimer's disease, Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, osteoporosis, arthritis, insulin-resistance and type II diabetes, benign prostatic hyperplasia, systemic lupus erythematosus, auto-immune diseases, psoriasis, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g, for prevention or as adjuvant therapy), eczema, pain, inflammation or strain of muscle, tendon or joint, lipolysis (e.g., by cold, heat, laser, ultrasound, or radiofrequency ablation for fat reduction and modification as a cosmetic procedure or a treatment for obesity), post lipectoniy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, arthritis (such as osteoarthritis and

rheumatoid arthritis), and combinations thereof. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytie p53/pRb activator and/or the NAD+ supplement are administered systemically. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytie p53/pRb activator and/or the NAD+ supplement are administered locally .

[0056] In some embodiments, there is provided a method of treating or preventing an aging- related disease or condition in an individual (e.g., a human or a pet animal), comprising administering to the individual: (a) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (b) an effective amount of a senolytie p53/pRb activator; and (c) an effective amount of a NAD+ supplement. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin. In some embodiments, the senolytie p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytie p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. In some embodiment, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytie p53/pKb activator and the NAD+ supplement are formulated in a single

pharmaceutical composition. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytie p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the aging-related disease or condition is selected from the group consisting of cancer (e.g, for prevention or as adjuvant therapy), cardiovascular diseases, central nervous system diseases, Alzheimer's disease, Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, osteoporosis, arthritis, insulin-resistance and type II diabetes, benign prostatic hyperplasia, systemic lupus erythematosus, auto-immune diseases, psoriasis, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g, for prevention or as adjuvant therapy), eczema, pain, inflammation or strain of muscle, tendon or joint, lipolysis (e.g., by cold, heat, laser, ultrasound, or radiofrequency ablation for fat reduction and modification as a cosmetic procedure or a treatment for obesity), post lipectomy and liposuction, non-alcoholic fatty liver disease, non alcoholic steatotic hepatitis, arthritis (such as osteoarthritis and rheumatoid arthritis), and combinations thereof. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are administered systemically. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are administered locally.

[0057] In some embodiments, there is provided a method of treating or preventing an aging- related disease or condition in an individual, comprising locally administering to the individual: (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nulraceutica! agent. In some embodiments, the mTOR inhibitor comprises rapamyein or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some

embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor (e.g., torin 1, torin 2, PP242 or INK! 28). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolvtic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the aging-related disease or condition is selected from the group consisting of primary ' or metastatic cancer lesions (e.g., for prevention or as adjuvant therapy), cardiovascular diseases, central nervous system diseases, Alzheimer's disease,

Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g., for prevention or as adjuvant therapy), psoriasis, eczema, pain, inflammation or strain of muscle, tendon or joint, obesity, cosmetic lipolysis, post lipectomy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatosis hepatitis, arthritis (such as osteoarthritis and rheumatoid arthritis), auto-immune diseases, and combinations thereof. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pKb activator, and/or the NAD+ supplement is administered via an implanted device. In some embodiments, the implanted device is selected from the group consisting of an intravesical or other urogenital tract gel, an intra-cranial or intra-thecal device, intra-ocular device, a drug-eluting stent, and CT, MRI or X-ray guided and inserted devices.

[0058] In some embodiments, there is provided a method of treating or preventing an aging- related disease or condition in an individual, comprising administering to the individual: (a) an effective amount of metformin or an insulin sensitizing nutraceutical agent; (b) an effective amount of ibuprofen or a rapamycin-iike nutraceutical agent; (c) an effective amount of rapamycin; and (d) an effective amount of niacin and an effective amount of nicotinamide riboside. In some embodiments, the metformin or an insulin sensitizing nutraceutical agent, ibuprofen, rapamycin or a rapamycin-like nutraceutical agent, NA and NR are administered orally. In some embodiments, the metformin is administered once, twice or three times daily after rneal(s). In some embodiments, metformin is administered at about 250 mg/dose to about 1000 mg/dose (e.g., about 250 mg/dose, about 500 mg/dose, about 750 mg/dose, or about 1000 mg/dose). In some embodiments, the ibuprofen is administered twice daily for two days per week, or twice daily for 4 weeks out of 6 weeks. In some embodiments, ibuprofen is

administered at about 400 mg/dose to about 600 mg/dose (e.g., about 400 mg/dose or about 600 mg/dose). In some embodiments, rapamycin or a rapamycin-like nutraceutical agent is administered weekly or bi-weekly, 6 weeks out of every seven or eight weeks. In some embodiments, rapamycin is administered at about 1 g to 10 mg weekly (e.g., about any one of 1 mg, 2mg, 3 mg, 4 mg, 5 mg, 5mg or 10 mg weekly). In some embodiments, NA is administered twice daily. In some embodiments, NA is administered at about 125 mg/dose to about 2000 mg/dose (e.g, about 125 mg/dose, about 250 mg/dose, or about 2000 mg/dose). In some embodiments, NR is administered once twice or four times daily. In some embodiments, NR is administered at about 125 mg/dose to about 1000 mg/dose (e.g., about 250 mg/dose, about 500 mg/dose, or about 1000 mg/dose).

[0059] In some embodiments, there is provided a method of treating or preventing an aging- related disease or condition in an individual, comprising administering to the individual: (a) an effective amount of metformin or an insulin sensitizing nutraceutical agent; (b) an effective amount of ibuprofen; (c) an effective amount of rapamycin or a rapamycin-!ike nutraceutieal agent and/or (d) an effective amount of a pan-mTOR inhibitor; and (e) an effective amount of niacin and an effective amount of nicotinamide riboside. In some embodiments, the pan-mTOR inhibitor comprises an agent having a low molecular weight (e.g., no more than about 500 Dalton). In some embodiments, the metformin, ibuprofen, rapamycin, NA and NR are

administered orally. In some embodiments, the metformin or an insulin sensitizing nutraceutieal agent is administered once, twice or three times daily after meal(s). In some embodiments, metformin is administered at about 250 mg/dose to about 1000 mg/dose (e.g., about 250 mg/dose, about 500 mg/dose, about 750 mg/dose, or about 1000 mg/dose) hi some

embodiments, the ibuprofen is administered twice daily for two days per week, or twice daily for 4 weeks out of 6 weeks. In some embodiments, ibuprofen is administered at about 400 mg/dose to about 600 mg/dose (e.g., about 400 mg/dose or about 600 mg/dose). In some embodiments, rapamycin or a rapamycin-like nutraceutieal agent is administered weekly, bi-weekly, once every' three weeks, weekly for 6 weeks out of 7 weeks, or weekly 6 weeks out of 8 weeks. In some embodiments, rapamycin is administered at about 1 mg to 10 mg weekly (e.g, about any one of 1 mg, 2mg, 3mg, 4 mg, 5 mg, 5mg or 10 mg weekly). In some embodiments, NA is administered twice daily. In some embodiments, NA is administered twice daily. In some embodiments, NA is administered at about 125 mg/dose to about 2000 mg/dose (e.g · ., about 125 mg/dose, about 250 mg/dose, or about 2000 mg/dose). In some embodiments, NR is

administered once, twice or four times daily. In some embodiments, NR is administered at about 125 mg/dose to about 1000 mg/dose (e.g · ., about 250 mg/dose, about 500 mg/dose, or about 1000 mg/dose). In some embodiments, the pan-mTOR inhibitor is administered topically.

In some embodiments, the pan-mTOR inhibitor is torin 1 , torin 2, PP242, or INK 128 In some embodiments, the aging related disease or condition is a skin disease or condition selected from the group consisting of seborrheic keratosis, actinic keratosis, hair loss and alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g., for prevention or as adjuvant therapy), skeletal muscular diseases, aging skin pigmentation, psoriasis, eczema, pain, inflammation or strain of muscle, tendon or joint, lipolysis (e.g., by cold, heat, laser, ultrasound, or radiofrequency ablation for fat reduction and modification as a cosmetic procedure or a treatment for obesity), post iipectomy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, arthritis (such as osteoarthritis and rheumatoid arthritis), auto-immune diseases and

combination s thereof. [0060] In some embodiments, there is provided a method of treating a local disease or condition in an individual, comprising locally administering to the individual (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., evero!imus). In some embodiments, the mTOR inhibitor comprises a pan-mTQR inhibitor (e.g., torin 1, torin 2, PP242 or INK128).

In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM.

In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the local administration is topical administration. In some embodiments, the local disease or condition is a skin disease or condition selected from seborrheic keratosis, actinic keratosis, hair loss and alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g., for prevention or as adjuvant therapy), obesity, cosmetic lipolysis, post lipectomy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatosis hepatitis, and combinations thereof. In some embodiments, the skin disease or condition is an allergic or auto-immune skin disease, such as eczema or psoriasis. In some embodiments, the local disease or condition is a disease of the muscular skeletal system, including but not limited to ail forms of arthritis such as

osteoarthritis, rheumatoid arthritis, auto-immune diseases, tendon and muscle pain and inflammation, and combinations thereof. In some embodiments, the method further comprises locally administering an effective amount of a corticosteroid. In some embodiments, the method further comprises locally administering an anti-oxidant, such as Chayote (Sechium edule) juice and/or extract. In some embodiments, the local administration is a guided intra-lesion or intra- regional visceral treatment. I some embodiments, the local disease or condition is a visceral disease or condition such as primary or metastatic cancer lesions (e.g., for prevention or as adjuvant therapy), post cancer resection area as an adjuvant for the prevention of future relapse, cardiovascular diseases, central nervous system diseases, Alzheimer’s disease, Parkinson's disease, age-related macular degeneration, cataract, or retinopathy.

[0061] In some embodiments, there is provided a method of treating or preventing an aging- related disease or condition in an individual ( e.g , a human or a pet animal), comprising administering to the individual: (a) an effective amount of metformin; (b) an effective amount of diclofenac; and (c) an effective amount of NR. In some embodiment, metformin, diclofenac and NR are formulated in a single pharmaceutical composition. In some embodiments, metformin, diclofenac and NR are formulated in two or more pharmaceutical compositions. In some embodiments, the aging-related disease or condition is selected from the group consisting of cancer (e.g., for prevention or as adjuvant therapy), cardiovascular diseases, central nervous system diseases, Alzheimer's disease, Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, osteoporosis, arthritis, insulin-resistance and type II diabetes, benign prostatic hyperplasia, systemic lupus erythematosus, auto-immune diseases, psoriasis, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g., for prevention or as adjuvant therapy), eczema, pain, inflammation or strain of muscle, tendon or joint, lipolysis (e.g., by cold, heat, laser, ultrasound, or radiofrequency ablation for fat reduction and modification as a cosmetic procedure or a treatment for obesity), post lipectomy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, arthritis (such as osteoarthritis and rheumatoid arthritis), and combinations thereof. In some embodiments, metformin, diclofenac and NR are administered systemieally. In some embodiments, metformin, diclofenac and NR are administered locally. In some embodiments, the method further comprises administering to the individual an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, 1,- carnitine, R-lipoic acid, ieptine, a caspase inhibitor, and/or Chayote (Sechinm edule) juice or extract. In some embodiments, the method comprises administering to the individual a pharmaceutical composition comprising about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) NR, about 0.5-6% (w/v) NA, and about 0.5-10% (w/v) pterostilbene. In some embodiments, the pharmaceutical composition further comprises about 1- 10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40% (w/v) PEG200, and about 5-40% (w/v) PEG400. In some embodiments, the pharmaceutical composition is administered to the individual topically. In some embodiments, the pharmaceutical composition comprises about 2% (w/v) diclofenac, about 2% (w/v) metformin, about 3% (w/v) NR, about 1.5% (w/v) NA, and about 1.5% (w/v) pterostilbene.

[0062] In some embodiments, there is provided a method of providing cosmetic treatment of skin in an individual, comprising topically administering to the skin of the individual: (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTQR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g, piogiitazone) and/or a higuanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapa!og (e.g., everolimus). In some

embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor (e.g., torin 1, torin 2, PP242 or INK 128). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g, ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the method is used for treating aging skin pigmentation and/or wrinkles. In some embodiments, the method further comprises locally administering an anti-oxidant, such as Chayote (Sechium edule) juice and/or extract.

[0063] In some embodiments, there is provided a method of providing cosmetic treatment of skin in an individual, comprising topically administering to the skin of the individual: (a) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (b) an effective amount of a senolytic p53/pRb activator; and (c) an effective amount of a NAD+ supplement. In some embodiments, there is provided a method of providing cosmetic treatment of skin in an individual, comprising topically administering to the skin of the individual: (a) an effective amount of metformin; (b) an effective amount of diclofenac; and (c) an effective amount of NR. In some embodiment, metformin, diclofenac and NR are formulated in a single pharmaceutical composition. In some embodiments, the method further comprises administering to the individual an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-camitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Chayote ( Sechium edule) juice or extract. In some embodiments, the method comprises administering to the individual a pharmaceutical composition comprising about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) NR, about 0.5-6% (w/v) NA, and about 0.5-10% (w/v) pterostilbene. In some embodiments, the pharmaceutical composition further comprises about 1-10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5- 40% (w/v) PEG200, and about 5-40% (w/v) PEG400. In some embodiments, the pharmaceutical composition comprises about 2% (w/v) diclofenac, about 2% (w/v) metformin, about 3% (w/v) NR, about 1.5% (w/v) NA, and about 1.5% (w/v) pterostilbene.

[0064] In some embodiments, there is provide a method of preventing, inhibiting, or reversing one or more signs of aging in a cell (e.g., mammalian cell) comprises contacting the cel! with (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (d) an effective amount of a N AD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some

embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor (e.g., torin 2). In some embodiments, the senolytic p53/pRb activator is selected from a nutiin or analog thereof, a COX inhibitor (e.g., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof.

In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some

embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions.

[0065] In some embodiments, there is provide a method of preventing, inhibiting, or reversing one or more signs of aging in a cel! (e.g., mammalian ceil) comprises contacting the cel! with: (a) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (b) an effective amount of a senolytic p53/pRb activator; and (c) an effective amount of a NAD+ supplement. In some embodiments, the agent having dual functions as an insulin- sensitizer and an mTOR inhibitor is metformin. In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a

CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD-ί- supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. In some embodiment, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the method further comprises contacting the cell with an effective amount of vitamin D, vitamin E, vitamin A, resveratroi, pterostilbene, L-carnitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Chayote ( Sechium edule) juice or extract.

[0066] In some embodiments, there is provided a method of stabilizing, decelerating and/or ameliorating one or more signs of aging in a plurality of mammalian cells by contacting the plurality of mammalian cells with (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g, pioglitazone) and/or a biguanide (e.g, metformin) hi some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor (e.g, torin 2). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g·., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the plurality of mammalian cells is present in a human. In some embodiments, the human has not previously been administered with the composition. In another embodiment, the human has previously been administered one, two or three regimen selected from the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD-ί- supplement in some embodiments, the plurality of mammalian cells becomes free of quiescent or senescent phenotypes after the contacting. In some embodiments, prior to the contacting, the plurality of mammalian cells is quiescent. In some embodiments, prior to the contacting, the plurality of mammalian cells is senescent. In some embodiments, prior to the contacting, the plurality of mammalian cells comprises both quiescent and senescent cells.

[0067] In some embodiments, there is provided a method of stabilizing, decelerating and/or ameliorating one or more signs of aging in a plurality of mammalian cells by contacting the plurality of mammalian cells with: (a) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (b) an effective amount of a senolytic p53/pRb activator; and (c) an effective amount of a NAD+ supplement. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin. In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g, diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. In some embodiment, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single

pharmaceutical composition. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the plurality of mammalian cells is present in a human. In some embodiments, the human has not previously been administered with the composition. In another embodiment, the human has previously been administered one, two or three regimen selected from the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement. In some embodiments, the method further comprises contacting the plurality of mammalian cells with an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-carnitine, R~ lipoic acid, leptine, a caspase inhibitor, and/or Chayote ( Sec ium edule) juice or extract. In some embodiments, the plurality of mammalian cells becomes tree of quiescent or senescent phenotypes after the contacting. In some embodiments, prior to the contacting, the plurality of mammalian cells is quiescent. In some embodiments, prior to the contacting, the plurality of mammalian cells is senescent. In some embodiments, prior to the contacting, the plurality of mammalian cells comprises both quiescent and senescent cells.

[0068] Signs of aging in cells ( .e., cellular aging) include, but are not limited to, shortening of telomeres, cell cycle arrest, senescence, mitochondrial dysfunction, deregulated cellular signaling (e.g., rnTOR, p53 and insulin signaling), change in metabolic state (e.g, to RIG state, i.e., Reproduction, Immune-aetivati on/Inflammation and Growth), altered level of protein synthesis, accumulation of DNA damage, and genome instability. Signs of cellular aging can be assessed using known methods in the art, including, but not limited to, ELISA, Western blot, RT-PCR, microscopy, in situ hybridization, DNA sequencing, etc. Such methods can be used to measure, for example, telomere length, expression levels of cytokines {e.g, 1L6, IL17), cell aging hypertrophy, protein and fat accumulation, etc.

[0069] In some embodiments, there is provided a method of converting a mammalian cell from senescent state to quiescent state or normal state, comprising contacting the mammalian cell with (a) an effective amount of an insulin sensitizer, (b) an effective amount of an rnTOR inhibitor, (c) an effective amount of a senolytic p53/pRb activator, and (cl) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g·., piogiitazone) and/or a biguanide {e.g, metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceuticai agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceuticai agent, and/or a rapalog {e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor {e.g, torin 2). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g·., ihuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM.

In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions.

[0070] In some embodiments, there is provided a method of converting a mammalian cell from senescent state to quiescent state or normal state, comprising contacting the mammalian ceil with (a) an effective amount of an agent having dual functions as an insulin sensitizer and an rnTOR inhibitor; (b) an effective amount of a senolytic p53/pRb activator; and (c) an effective amount of a NAD+ supplement. In some embodiments, the agent having dual functions as an insulin-sensitizer and an niTOR inhibitor is metformin in some embodiments, the senoiytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senoiytic p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. In some embodiment, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senoiytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTO inhibitor, the senoiytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the method further comprises contacting the cell with an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-carnitine, R-lipoic acid, lepline, a caspase inhibitor, and/or Chayote ( Sechium edule) juice or extract.

[0071] In some embodiments, the method is applicable for prophylactic and/or therapeutic treatment of symptoms and functional abnormalities of aging ceils having the senescence associated secretory phenotype (SASP). SASP cells contributes to aging-related diseases such as Alzheimer’s disease, cardiovascular system atherosclerosis which can lead to stroke, heart attack and heart failure, prostate enlargement, cardiac hypertrophy, arterial wall thickening, osteoarthritis, age-related macular degeneration, etc. SASP cells also secrete IL6 The combination regimen described herein can decrease the number of SASP ceils, and convert them to a quiescence or normal phenotype, as well as reducing the IL6 level in an individual.

[0072] In some embodiments, there is provided a method of inhibiting or reducing SASP cells in an individual in need thereof, comprising administering to the individual: (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senoiytic p53/pRb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan -mTOR inhibitor (e.g., torin 2). In some embodiments, the seno!ytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g, ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD-- supplement are formulated in a single phar aceutical composition. In some

embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and/or the NAIH supplement are administered systemically. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are administered locally.

[0073] In some embodiments, there is provided a method of inhibiting or reducing SASP cells in an individual in need thereof, comprising administering to the individual: (a) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (b) an effective amount of a senolytic p53/pRb activator; and (c) an effective amount of a NAD supplement. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin. In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g , diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. In some embodiment, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the method further comprises administering to the individual an effective amount of vitamin D, vitamin E, vitamin A, resveratroi, pterostilbene, L- camitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Chayote (Sechium edule) juice or extract. [0074] In some embodiments, there is provided a method of reducing IL-6 level in an individual in need thereof, comprising administering to the individual: (a) an effective amount of an insulin sensitizer, (b) an effective amount of an mTOR inhibitor, (c) an effective amount of a senolytic p53/pKb activator, and (d) an effective amount of a NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the rnTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan-mTGR inhibitor (e.g., torin 2). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g, ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiment, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and the NAD supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. I some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are administered systemically. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD supplement are administered locally.

[0075] In some embodiments, there is provided a method of reducing IL-6 level in an individual in need thereof, comprising administering to the individual : (a) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (b) an effective amount of a senolytic p53/pRb activator, and (c) an effective amount of a NAD · supplement. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin hi some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises nicotinic acid (NA), nicotinamide riboside (NR), nicotinamide mononucleotide (NMN), and/or nicotinamide (NAM). In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. In some embodiment, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions in some embodiments, the method further comprises administering to the individual an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-camitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Chayote ( Sechium edule) juice or extract.

[0076] In some embodiments, the insulin sensitizer is any one or more agents listed in the section“A. Insulin sensitizer” below, including any combinations thereof The various agents can be administered via the same or different routes of administration, and in any suitable order. In some embodiments, the insulin sensitizer comprises an agent of the thiazolidinedione (TZD) family, such as pioglitazone, or rosiglitazone. In some embodiments, the insulin sensitizer comprises a biguanide, such as metformin. In some embodiments, the insulin sensitizer comprises an insulin sensitizing nutraeeutical agent.

[0077] In some embodiments, the insulin sensitizer comprises a thiazolidinedione (TZD) and/or a biguanide. In some embodiments, the insulin sensitizer is metformin. In some embodiments, the insulin sensitizer comprises an insulin sensitizing nutraeeutical agent. In some embodiments, the insulin sensitizer is a combination of TZD (e.g, pioglitazone) and metformin. In some embodiments, the insulin sensitizer is administered orally. In some embodiments, the insulin sensitizer is administered once daily, twice daily, or three times daily in some embodiments, the insulin sensitizer is administered after meals. In some embodiments, metformin is administered at a dose of no more than about 3000 mg per day, such as no more than about any one of 750 mg, 1000 mg, 1250mg, ISOOmg, 2Q00mg, 2500mg or 3,000 mg per day. In some embodiments, metformin is administered at a dose of about 250 mg/dose to about 1000 mg/dose, such as about any one of 250 mg/dose, 500 mg/dose, 750 mg/dose, or 1000 mg/dose.

[0078] In some embodiments, the mTOR inhibitor is any one or more agents listed in the section“B. mTOR inhibitor” below, including any combinations thereof. The various agents can be administered via the same or different routes of administration, and in any suitable order. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraeeutical agent, and/or a rapalog. In some embodiments, the rapalog is selected from the group consisting of everolimus, tacrolimus, CCI-779, ABT-578, AP -23675, AP-23573, AP -23841, 7-epi- rapamycin, 7-thiomethyl -rapamycin, 7-epi-trimethoxyphenyl -rapamycin, 7-epi-thiomethyl- rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-rapamycin, 7-demethoxy- rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin, 42-0-(2-hydroxy)ethyi rapamycin, and combinations thereof. In some embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor, also known as“dual mTORCl/TORC2 inhibitor” (i.e., an inhibitor that inhibits both TORC1 and TORC2). In some embodiments, the pan-mTOR inhibitor is selected from the group consisting of sapanisertib (GNK128), AZD2014, torin 1, torin 2, AZD8055,

PP242, KU-006379, OSI-027, WAY-600, WAE-687, WYE-354 and GSK 1059615. In some embodiments, the mTOR inhibitor comprises an indirect AMPK activator. In some embodiments, the mTOR inhibitor comprises a direct AMPK activator. In some embodiments, the AMPK activator is metformin or an insulin sensitizing nutraceutical agent or any agents in the biguanide group. In some embodiments, the mTOR inhibitor comprises an agent that inhibits PI3K and/or AKT. In some embodiments, the mTOR inhibitor comprises any combination of: (a) a

rapamycin or a rapamycin-like nutraceutical agent and/or a rapalog; (b) a pan-mTOR inhibitor;

(c) a direct AMPK activator and/or an indirect AMPK activator; and/or (d) an agent that inhibits PI3K and/or AKT.

[0079] In some embodiments, the mTOR inhibitor is rapamycin or a rapamycin-like

nutraceutical agent. In some embodiments, the mTOR inhibitor is everolimus. In some embodiments, the mTOR inhibitor is administered orally. In some embodiments, the mTOR inhibitor is administered weekly. In some embodiments, the mTOR inhibitor is administered bi weekly. In some embodiments, the mTOR inhibitor is administered in cycles of 6 weeks on and 1 week off. In some embodiments, the mTOR inhibitor is administered in cycles of 6 weeks on and 2 weeks off. In some embodiments, the effective amount of rapamycin or the rapalog maintains a trough blood level of rapamycin selected from no more than about 8ng/ml for systemic administration, no more than about 2ng/ml for local administration, and no more than about lOng/mi for combined systemic and local administration, or an equivalent trough blood level of the rapalog thereof In some embodiments, rapamycin is administered at a dose of no more than about 10 mg/week, such as no more than about any one of 8mg/week, 6 mg/week, 3 mg/week, 2 mg/week, or 1 mg/week.

[0080] In some embodiments, the senolytic p53/pRb activator is any one or more agents listed in the section“C. senolytic p53/pRb activator” below, including any combinations thereof. The various agents can be administered via the same or different routes of administration, and in any suitable order. In some embodiments, the senolytic p53/pRb activator comprises an agent that activates or stabilizes p53, such as nutlin or analog thereof (e.g., nutlin-3A). In some embodiments, the senolytic p53/pRb activator comprises an agent that inhibits a Sirtuin, such as SirTI and SirT2. In some embodiments, the senolytic p53/pRb activator comprises a COX inhibitor, such as a COX2 inhibitor. In some embodiments, the COX inhibitor is selected from the group consisting of voltaren, aspirin, ibuprofen, naproxen, diclofenac, indometbacin, and combinations thereof. In some embodiments, the senolytic p53/pRb activator comprises an agent that activates or dephosphorylates pRb, such as a pan CDK inhibitor, or a CDK4/6 inhibitor (/.<?., an inhibitor that specifically inhibits CDK 4/C } In some embodiments, the pan CDK inhibitor is selected from the group consisting of tlavopiridol, olomoucine II, purvalanol A, SNS-032, dinaciclib, MK-7965, SCH727965, AT7519, R547, AZD5438, and AG024322. In some embodiments, the CDK4/6 inhibitor is selected from the group consisting of ribociclib, paiboeiclib, abemaciclib, and trilaciclib

[0081] In some embodiments, the senolytic p53/pKb activator is selected from a nutlin or analog thereof, a COX inhibitor, a CDK4/6 inhibitor, a pan-CD K inhibitor, and combinations thereof. In some embodiments, the senolytic p53/pKb activator is ibuprofen. In some

embodiments, the senolytic p53/pRb activator is administered orally. In some embodiments, ibuprofen is administered twice daily for two days per week, or twice daily for 4 weeks out of 6 weeks. In some embodiments, ibuprofen is administered at a dose of about 400 mg/dose to about 600 mg/dose, such as about any one of 400 mg, 450 mg, 50Qmg or 600 mg per dose.

[0082] In some embodiments, the NAD+ supplement is any one or more agents listed in the section“D. NAD+ supplement” below, including any combinations thereof. The various agents can be administered via the same or different routes of administration, and in any suitable order. In some embodiments, the NAD+ supplement comprises an agent from the de novo crossover pathway, such as Nicotinic Acid or Niacin (NA). In some embodiments, the NAD+ supplement comprises an agent from the salvage pathway, such as Nicotinamide Riboside (NR),

Nicotinamide mononucleotide (NMN), and/or Nicotinamide (NAM). In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NANI. In some embodiments, the NAD+ supplement comprises: (a) NA and (b) one or more of NR, NMN and NAM. In some

embodiments, the NAD+ supplement comprises NR and NA.

[0083] In some embodiments, the NAD+ supplement comprises NA and NR In some embodiments, the NAD+ supplement is administered orally. In some embodiments, the NAD+ supplement is administered twice daily. In some embodiments, the NA is administered at a dose of at. least about 125 mg per day, such as at least about any one of 150 mg, 200 mg, 250mg, 300 mg, 400 g, 500 mg, 1000 mg, 2000 mg, or 3000 mg per day. In some embodiments, the NR is administered at a. dose of at least about 250 mg per day, such as at least about any one of 400 mg, 500 mg, 600 mg, 800 mg, lOQOmg, or 2000 mg per day. In some embodiments, the effective amount of the NAD+ supplement maintains a blood level of at least about 500mM NAD+.

[0084] In some embodiments, any two of the insulin sensitizer, the mTOR inhibitor, the senolytic p53/pKb activator and the NAD+ supplement can comprise of, or being represented by, only one agent with dual component properties. In some embodiments, any one or more, or ail of the components in the combination regimen can comprise of, or being represented by

nutraceutical agents with the same component properties.

[0085] In some embodiments, the insulin sensitizer and the mTOR inhibitor, or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor; the senolytic p53/pRb activator; and the NAD+ supplement are administered simultaneously or sequentially in any suitable order. In some embodiments, each component of the combination regimen, i.e., the insulin sensitizer, the mTOR inhibitor, or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor; the senolytic p53/pKb activator; and the NAD+ supplement etc., is formulated as separate pharmaceutical compositions. In some embodiments, two or more components of the combination regimen are formulated as a. single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor; the senolytic p53/pRb activator; and the NAIH supplement are formulated as a single pharmaceutical composition. In some embodiments, wherein each component of the combination regimen comprises two or more agents, the two or more agents are formulated as a single pharmaceutical composition or separate pharmaceutical compositions.

[0086] In some embodiments, there is provided a pharmaceutical composition comprising two or more of: (i) an insulin sensitizer; (ii) an mTOR inhibitor; (iii) a senolytic p53/pRb activator; (iv) an NAD+ supplement, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is a pill, a tablet, a mixture, a solution, a cream, a liniment, an eye drop, or a nanoparticle composition.

[0087] In some embodiments, there is provided a pharmaceutical composition comprising two or more of: (i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor; (ii) a senolytic p53/pRb activator; (iii) an NAD+ supplement, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is a pill, a tablet, a mixture, a solution, a cream, a liniment, an eye drop, or a nanoparticle composition.

[0088] In some embodiments, there is provided a pharmaceutical composition comprising metformin, diclofenac, NR and/or NA, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises pterostilbene. In some embodiments, the pharmaceutical composition further comprises urea, sodium acetate, ethanol, PEG200, and/or PEG400. In some embodiments, the pharmaceutical composition is formulated for topical administration. In some embodiments, the pharmaceutical composition is a solution or a cream. In some embodiments, the pharmaceutical composition comprises about 0.5-10% (w/v) metformin, such as at least about any one of 0.5, 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10%, or no more than about any one of 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.5%, or about any one of 0.5-1%, 1-2%, 2- 5%, 5-10%, 0.5-2.5%, 2,5-5%, 5-5.75%, or 7.5-10% (w/v) metformin. In some embodiments, the pharmaceutical composition comprises about 0.5-4% (w/v) diclofenac, such as at least about any one of 0.5, 1, 1.5, 2, 2.5, 3, 3.5, or 4%, or no more than about any one of 4, 3.5, 3, 2.5, 2, 1.5, 1, or 0.5%, or about any one of 0.5-1%, 1 -2%, 2-3%, 3-4%, 0.5-2%, 2-4%, or 1-3% (w/v) diclofenac. In some embodiments, the pharmaceutical composition comprises about 2% (w/v) diclofenac. In some embodiments, the pharmaceutical composition comprises about 0.5-6%

(w/v) NR, such as at least about any one of 0.5, I, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5 or 6%, or no more than about any one of 6, 5.5, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.5, 1, or 0.5%, or about any one of 0.5-1, 1-2, 2-4, 4-6, 0.5-2, 1-4, 2-5, or 3-6% (w/v) NR. hi some embodiments, the

pharmaceutical composition comprises about 3% > (w/v) NR. In some embodiments, the pharmaceutical composition comprises about 0.5-6% (w/v) NA, such as at least about any one of 0.5, 1 , 1 5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5 or 6%, or no more than about any one of 6, 5.5, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.5, 1, or 0.5%, or about any one of 0.5-1, 1-2, 2-4, 4-6, 0.5-2, 1-4, 2-5, or 3-6% (w/v) NA. In some embodiments, the pharmaceutical composition comprises about 1.5% (w/v) NA. In some embodiments, the pharmaceutical composition comprises about 1 -10% (w/v) urea, such as at least about any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%, or no more than about any one of 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1%, or about any one of 1-2.5, 2.5-5, 5-7.5, 7.5-10, 1-5, 5-10, 2-4, 4-6, 6-8, or 8-10% (w/v) urea. In some embodiments, the pharmaceutical composition comprises about 1-10% (w/v) sodium acetate, such as at least about any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10% , or no more than about any one of 10, 9, 8, 7, 6, 5, 4, 3, 2, or l%o, or about any one of 1-2.5, 2.5-5, 5-7.5, 7.5-10, 1-5, 5-10, 2-4, 4-6, 6-8, or 8-10% (w/v) sodium acetate. In some embodiments, the pharmaceutical composition comprises about 1-30% (w/v) ethanol, such as at least about any one of 1, 2, 5, 10, 15, 20, 25, or 30%, or no more than about any one of 30, 25,

20, 15, 10, 5, 2, or 1%, or about any one of 1-5, 5-10, 10-15, 15-20, 20-25, 25-30, 1-10, 10-20, or 20-30% (w/v) ethanol. In some embodiments, the pharmaceutical composition comprises about 5-40% (w/v) PEG200, such as at least about any one of 5, 10, 15, 20, 25, 30, 35, or 40%, or no more than about any one of 40, 35, 30, 25, 20, 15, 10 or 5%, or about any one of 5-10, 10- 15, 15-20, 20-30, 30-40, 5-20, 20-40, or 10-30% (w/v) PEG200. In some embodiments, the pharmaceutical composition comprises about 5-40% (w/v) PEG400, such as at least about any one of 5, 10, 15, 20, 25, 30, 35, or 40%, or no more than about any one of 40, 35, 30, 25, 20, 15,

10 or 5%, or about any one of 5-10, 10-15, 15-20, 20-30, 30-40, 5-20, 20-40, or 10-30% (w/v) PEG400. In some embodiments, the pharmaceutical composition further comprises about 0.5-10% (w/v) pterostilbene, such as at least about any one of 0.5, I, 2, 3, 4, 5, 6, 7, 8, 9, or 10%, or no more than about any one of 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.5%, or about any one of 0.5-1%, 1 - 2%, 2-3%, 3-5%, 5-10%, 0.5-5%, 2.5%-7.5%, 1-3%, 3-6% or 6-9% (w/v) pterostilbene. In some embodiments, the pharmaceutical composition comprises about 1.5% (w/v) pterostilbene. In some embodiments, the pharmaceutical composition further comprises water.

[0089] In some embodiments, there is provided a pharmaceutical composition comprising about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) NR, about 0.5-6% (w/v) NA, about 0.5-10% (w/v) pterostilbene, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises urea, sodium acetate, ethanol, PEG200, and/or PEG400. In some embodiments, the pharmaceutical composition comprises about 2% (w/v) diclofenac, about 2% (w/v) metformin, about 3% (w/v)

NR, about 1.5% NA, and about 1.5% (w/v) pterostilbene. In some embodiments, the

pharmaceutical composition is formulated for topical administration. In some embodiments, the pharmaceutical composition is a solution or a cream.

[0090] In some embodiments, there is provided a pharmaceutical composition comprising about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) NR, about 0.5-6% (w/v) NA, about 0.5-10% (w/v) pterostilbene, about 1-10% (w/v) urea, about 1-10%

(w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40% (w/v) PEG20Q, and about 5-40% (w/v) PEG400.

[0091] The insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and the NAD+ supplement can be administered to the individual via any suitable route of administration. The methods described herein further contemplates in vitro or ex vivo use of the combination regimen. One of skill in the art. will recognize that the form and character of the particular dosing regimen employed in the methods of the present invention depend on the route of administration and other well-known variables, such as rate of clearance, the size of the individual, the stage of the particular disease being treated, toxicities, rate and total penetration enhancement of the medium and excipient, etc. Based on such criteria, one skilled in the art can determine an effective amount of any of the particular compositions described herein that will be effective for prophylaxis and/or therapy of aging-related diseases and/or for cellular aging in an individual.

[0092] in some embodiments, the individual is a human individual. In some embodiments, the human individual is at least about 50 years old, such as at least about any one of 55, 60, 65, 70, 75, 80 years old or higher. In some embodiments, the individual is non-diabetic. In some embodiments, the individual has impaired glucose regulation (IGR) or pre-diabetic. In some embodiments, the individual has diabetes, such as type diabetes. In some embodiments, the individual has an HbAlC level of no more than 42 mmol/mol or no more than 6.0%. In some embodiments, the individual has an HbAl C level of about 42 to about 47 mmol/mol, or about 6.0-6.4%. in some embodiments, the individual has an HbAlC level of higher than 48

mmol/mol or 6.5% or higher. HbAlC levels in a blood sample from the individual can be measured using commercially available kits. Together with the fasting plasma glucose test, the HbAlC test can be used to assess insulin resistance/sensitivity and status of diabetes in an individual.

[0093] In some embodiments, the individual is a pet animal or a sport animal, such as a cat, a dog, or a horse. In some embodiments, the dosages of the insulin sensitizer, the rnTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytie p53/pRb activator, and the NAD+ supplement are adjusted according to the average body weight, body surface area and pharmacological parameters (e.g, clearance and metabolism) specific to the individual’s species. In some embodiments, the effective amount of an agent for a pet (e.g., dog or cat) is no less than about 20% (such as about any one of 10%, 5%, 2%, or 1%) of the effective amount of the agent for a human individual. In some embodiments, the effective amount of metformin for a pet animal is about 5 mg twice daily, or about 7.5 mg twice daily. In some embodiments, the effective amount of ibuprofen for a pet animal is about 4 mg twice daily for 2 days/week, or 6 mg daily for 2 days/week. In some embodiments, the effective amount of rapamycin for a pet animal is about IQpg weekly. In some embodiments, the effective amount of the NA for a pet animal is about 1.25 mg twice daily or about 2.5 mg twice daily. In some embodiments, the effective amount of the NR for a pet animal is about 2.5 mg twice daily or about 5 mg twice daily.

[0094] The insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the sen y tic p53/pRb activator, and the NAD+ supplement can be administered via any suitable routes of administration, and/or via any suitable device. The agents of the combination regimen can be administered via the same routes or different routes, depending on the nature of the aging-related disease or condition being treated or prevented. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered via routes that are most amenable for long-term administration, e.g, by gastrointestinal routes or topical route in some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered via routes that reduce or minimize toxicity of the agents to the individual. For example, agents with narrow therapeutic window, such as highly potent mTOR inhibitors and senolytic p53 activators, may be administered locally. Suitable routes, formulations, and devices for administration of the combination regimen are described in the sections“E. Administration” and“F. Indications.”

[0095] In some embodiments, one or more of the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and the NAD+ supplement is administered locally. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered topically. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered via an implanted device in a tissue or organ.

[0096] In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered systemieally. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered orally. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are administered via a combination of systemic and local routes. For example, a first combination regimen or component thereof is administered to the individual locally, and a second combination regimen or component thereof is administered systemieally prior to, concurrently with, or subsequent to the first combination regimen or component thereof.

[0097] Any of the methods and compositions described herein may be administered in conjunction with another conventional therapy for treating or preventing an aging-related disease or condition, and/or cellular aging. For example, in the case of cancers, conventional therapies include, but are not limited to, chemotherapies, radiation therapy and surgical interventions. For skin diseases or conditions, corticosteroid is an exemplary conventional therapy. Additional anti- aging compounds or agents that augment, supplement, or synergize with the combination regimen can be administered in conjunction to the combination regimen to the individual.

Suitable anti-aging compounds include, but are not limited to, vitamins and over-the-counter supplements such as vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-earnitine, R- lipoic acid, leptine, caspase inhibitors, and/or antioxidant compounds {e.g., Chayote (Sechium edule) juice or extract) and combinations thereof.

[0098] In some embodiments, one or more antioxidant compounds are administered to the individual in conjunction to the combination regimen. The antioxidant compounds can be natural or man-made, and they can be derived from plants, vegetables, fruits and their juices and/or extracts and combinations thereof. In some embodiments, the antioxidant compound is

Chayote (Sechium edule) juice and/or extract. In some embodiments, the antioxidant compound is a polyphenol, a lycopene, a xanthine, a flavonoid, a derivative or an analog thereof.

[0099] In some embodiments, one or more anti-inflammatory agents are administered in conjunction to the combination regimen to the individual. Suitable anti-inflammatory' agents include, but are not limited to, !eukotrienes inhibitors, HMCo-A reductase inhibitors such as statins, antibodies against pro-inflammatory cytokines, chemokines and combinations thereof.

[0100] In some embodiments, a cell therapy is administered in conjunction to the combination regimen to the individual. Exemplary ' cell therapies include, but are not limited to,

administration of autologous and/or allogeneic somatic cells {e.g., neurons and immune cells), or autologous and/or allogeneic toti~, multi- or pluripotent stem cells. The stem cells can be non- induced, or induced by genetic manipulation (e.g., iPS by KOSM, Klf4, Oct3/4, Sox2 and cMyc), by chemical or small molecule manipulation such as HDAC inhibitors, or by a chemical cocktail VCRFSGY (HDAC inhibitors such as valproic acid; CHIR99021; Repsox; SP600125 (INK inhibitor), G06983 (PKC inhibitor) and Y-27632 (ROCK inhibitor)). Neurons can be obtained by inducing stem cells with LDN193189, SB431542, TTNPB, Thiazovivin, CHIR99021, VP A,

N - [N -(3 , 5 -dill uorophenacetyl)-L-al anyl ]-S~ph enylglyci ne t-butyl ester (D APT), Smoothened agonist (SAG), and purmorphamine.

[0101] In some embodiments, a hormone is administered in conjugation to the combination regimen to the individual. Suitable hormones include, but are not limited to, hGH, testosterone, estrogens, progesterone, GnRH, gonadal stimulating hormones, corticosteroid, and combination thereof.

A. Insulin sensitizer

[0102] The methods described herein comprise administering an effective amount of an insulin sensitizer, which may comprise one or more insulin sensitizing agents.

[0103] Many insulin sensitizing agents are known in the art. In some embodiments, the insulin sensitizer comprises a biguanide such as Metformin, or an insulin sensitizing nutraceutical agent. Metformin is a well-known anti-diabetic drug used clinically for over fifty years, and its main mechanism of action is insulin sensitization.

[0104] In some embodiments, the insulin sensitizer comprises a direct PPARy agonist. PPARy agonists include, but are not limited to, thiazolidinediones (TZDs) and structurally diversified TZDs, PPARcc/y dual agonists, and PPAR pan agonist. Many PPARy agonists are chemical derivatives of the parent compound thiazolidinedione. Several PPARy agonists have been approved for clinical use for various indications. For example, pioglitazone (e.g., ACTOS } has been approved but with a warning on a possible bladder cancer risk. Rosiglitazone (e.g.,

AVANDIA * , which was put under selling restrictions in the US and withdrawn from the market in Europe due to an increased risk of cardiovascular events suggested by some studies. Upon re- evaluation of new data in 2013, the FDA lifted the restrictions. Lobeglitazone (e.g · ., DUVIE 5 ') was approved for use in Korea. Darglitazone, cig!itazone, englitazone, netoglitazone,

nivoglitazone, and troglitazone have also been approved. In some embodiments, the insulin sensitizer comprises a selective PPARy modulator, for example, a PPARy modulator that hinds to PPARy with no agonism and inhibits pSer273PPARy. [0105] In some embodiments, the insulin sensitizer comprises an angiotensin inhibitor, or a related agent thereof, such as an angiotensin converting enzyme (ACE) inhibitor, and an angiotensin receptor blocker (ARB). Exemplary' clinically available angiotensin inhibitors include, but are not limited to, benazepril (e.g., LOTENSIN ® ), captopril (e.g, CAPOTEN' ® ), enalapril (e.g., VASOTEC ® , EPANED ® , or LEXXEL ® ), fosinopril (MONOPRIL ® ), lisinopril (e.g., PRINIVIL ® , ZESTRIL ® , or QBRELIS ® ), moexipril (e.g., UNIVA8C ® ), perindopril (e.g · ., ACEQN ® ), quinapril (e.g., ACCUPRIL ® ), ramipril (e.g, ALT ACE ® ), trandolapril (e.g.,

MAV1K ® ), azilsartan (e.g, ED ARBI ® ), candesartan (e.g, AT AC AND 4 '), eprosartan, irbesartan (e.g, AVAPRO ® ), losartan (e.g, COZAAR ® ), olmesartan (e.g, BENICAR ® ), telmisartan (e.g, M1CARDIS ® ), and valsartan (e.g, DIO VAN ® ).

[0106] In some embodiments, the insulin sensitizer comprises a PPARy sparing compound. In some embodiments, the PPARy sparing compound is a ligand or modulator of mTQT such as a ligand of mito-NEET (i.e., TT01001), or a ligand/modulator of m TOT (i.e., MSDC -0602/0160). In some embodiments, the PPARy sparing compound is an agent that targets one or more PPARy downstream effectors, such as adiponectin activator, FGF receptor targeted ligand and a derivative of FGF21 and/or FGF1 In some embodiments, the PPARy sparing compound is an agent that stimulates HSP and/or NOS, such as RGP-15. In some embodiments, the PPARy sparing compound is a biguanide such as metformin.

[0107] In some embodiments, the insulin sensitizer also functions as an mTOR inhibitor.

[0108] hi some embodiments, the insulin sensitizer is metformin, or an insulin sensitizing nutraceutical agent. Exemplary insulin sensitizing nutraceutical agent can be an agent or an extract of a mixture of agents from Galega officinalis, commonly known also as galega, goat’s- rue, French lilac or professor-weed and with ingredients such as guanidine, galegine, hydroxvgalegine, guanidine derivatives such as 4-hydroxygaIegine flavones, flavone glycosides, kaemferol, and quercetin. It can also be an agent, its precursors or derivatives, that activate PPARgamma, such as Flavonoids (Luteolin, Kaempferol, -Catechin, 2’-Hydroxychalcone, Biochanin A, Genistein, 6-Hydroxydaidzein, 6 -Hydroxyl-0-desmethylangolensin), Neolignans (Honokiol, Magnolol, Resveratrol, Pterostilbene, Amorphastilbol), Amorfrutins (Amorfrutins 1, 2, B), Falcarindiol, Deoxyelephantopin, Sargaquinoic acid, and Sargahydroquinoic acid.

[0109] In some embodiments, the insulin sensitizer can be one of the pde5 inhibitors such as viagra (sildenafil), or cialis (tadalafil) etc. B. mTOR inhibitor

[0110] The methods described herein comprise administering an effective amount of an mTOR inhibitor, which may comprise one or more mTOR inhibiting agents.

[0111] Without being bound by any theory or hypothesis, the mTOR inhibitor can reset the metabolic state in the body at rest or fasting to the default state, i.e., changing the metabolic state from RIG (i.e , Reproduction, Immune-activation/lnflammation and Growth) to RCH (i.e.,

Repair/Regeneration, Conservation and Homeostasis). Other effects may accompany this change in metabolic state, including increased availability of NAD+, which is required for effective DNA repair and mitochondrial biogenesis. This reset towards RCH during rest and fasting may working synergistically with the senolytic p53/pRb inhibition by reducing senescent cells and restoring them back to a quiescent state, which is amenable to DNA repair. In addition, this reset towards RCH may promote proliferation of adult stern cells (ASCs), and preserve non-senescent ASCs. As used herein,“RIG to RCH default reset” refers to reset of body metabolism at rest or fasting from a default of reproduction, immune-activation/inflamrnation and growth to a default consisting of repair/regeneration, conservation and homeostasis. Together, mTOR inhibition, which via RIG to RCH reset, can promote regeneration, homeostasis, natural cell death, and tissue replenishment.

[0112] Many mTOR inhibiting agents are known in the art. Exemplary mTOR inhibiting agents include, but are not limited to, rapamycin or a rapamycin-like nutraceuticaf agent, and rapalogs, pan-mTOR inhibitor, and direct or indirect AMPK activators. In some embodiments, the AMPK activator is metformin, an insulin sensitizing nutraceutical agent, or an agent in the biguanide group. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog. Suitable rapalogs include, but are not limited to, rapamycin (sirolimus), evero!imus, temsirolimus, ridaforolimus, tacrolimus, CCI-779, ABT-578, AT -23675, AP-23573, AP -23841, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi- trimethoxyphenyl -rapamycin, 7-epi-thiomethy! -rapamycin, 7-demethoxy-rapamycin, 3,2- demethoxy-rapamycin, 2-rapamycin, 7 -demethoxy-rapamy tin, 3 ,2-demethoxy-rapamy tin, 2- desmethyl-rapamycin, and 4,2-0-(2-hydroxy)ethyl rapamycin, and combinations thereof.

[0113] In some embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor, also known as a dual mTORC I/TO C2 inhibitor. Exemplary pan-mTOR inhibitors include, but are not limited to, sapanisertib (INK 128), AZD2014, torin 1, torin 2, AZD8055, PP242, KU-006379, QSI-027, WAY-600, WAE-687, WYE-354 and GSK 1059615 In some embodiments, the pan- mTOR inhibitors have a small molecular weight. For example, torin 1 , torin 2, PP242 and INK 128 are about 500 Dalton, and thus are suitable for topical delivery. Thus, such pan-mTOR inhibitors may have better pharmacological profile in certain applications (e.g., topical administration) than rapamycin or a rapamycin-like nutraceuticai agent for promoting re- proliferation of senescent cells, cell hypertrophy and/or oil retention

[0114] In some embodiments, the mTOR inhibitor comprises a direct AMPK activator, or an indirect AMPK activator. Suitable indirect AMPK activators include, but are not limited to, pterostilbene, quercetin, genistein, epigailocatechin gallate, berberine, curcurmin, ginsenoside Rbl, alpha-lipoic acid, and cryptotanshinone. Suitable direct AMPK activators include, but are not limited to, 5-aminoimidazo!e-4-carboxamide riboside (i.e., AICAR), thienopyridone (e.g., A- 7.69662), benzimidazole (e.g.. Compound 911), salicylate or pro-drug of aspirin, Compound- 13 (i.e., Pro-drug C2), PT-1, and MT63-78 (i.e., Debio0930). Other AMPK activators have been described, for example, see WO2009124636, W02009100130, WO2011029855,

WO201 1138307, WO2011080277, WO2011032320, and WO201 1033099.

[0115] In some embodiments, the mTOR inhibitor comprises an agent that inhibits PI3K and/or AKT. The PI3K/AKT pathway is upstream of mTOR. Agents that can inhibit PI3K, or AKT, or combination inhibition of PBK and AKT, with or without directly inhibiting mTOR, are effective mTOR inhibitors. In some embodiments, the mTOR inhibitor comprises a PI3K inhibitor, such as an inhibitor of PI3K alpha, delta, and/or gamma. Exemplary PI3K inhibitors include, but are not limited to, taselisib, perifosine, idelalisib, buparlisib (BKM120), duvelisib (IPI-145), alpelisib (BYL719), umbralisib, (TGR 1202), copanlisib (BAY 80-6946), PX-866, dactolisib, CUDC-907, ME-401 (a PI3K-Delta inhibitor), IPI-549 (a PI3K-gamma inhibitor),

SF1126, RP6530 (a Dual PI3K delta/gamma inhibitor), INK! 117 (a PI3K-alpha inhibitor in phase I trial), pictilisib (GDC-0941), XL147 (SAR245408), XI.765 (SAR245409), palomid 529, GSKIQ596I5, ZSTK474, PWT33597 (a dual PBK-alpha/mTOR inhibitor), NVP-BEZ235 (a dual P13K/AKT inhibitor), IC87114, TGI 00-1 15, CAL263, RP6503, PI-103 (a dual P13K- mTOR inhibitor), GNE-477 (PI3K-a!pha and mTOR inhibitor), and AEZS-136 (Erkl/2 inhibitor).

[0116] In some embodiments, the mTOR inhibitor comprises an AKT inhibitor. In some embodiments, the AKT inhibitor is an ATP competitive inhibitor selected from isoquinoline-5- sulfonamides (e.g., H-8, H-89, NL-71-101), azepane derivatives, aminofurazans (e.g., GSK 690693), heterocyclic rings (e.g., 7-azaindole, 6-phenylpurine derivatives, pyrrolo[2,3- djpyrimidine derivatives, CCT 128930, 3-aminopyrrolidine, anilinotriazole derivatives, spiroindoline derivatives, AZD5363, ipatasertib (GDC-0068, RG7440), A-674563, A- 443654), and phenylpyrazole derivatives ( e.g ., AT7867, AT13148). In some embodiments, the AKT inhibitor is a thiophenecarboxamide derivative (e.g., GSK 2141795). In some embodiments, the AKT inhibitor is an allosteric inhibitor, such as a 2,3-diphenylquinoxaline analogue or derivative, triazolo, naphthyridin-3(2/: )-one derivative (e.g., MK-2206); an alkylphospholipid such as ede!fosine (l-0-octadecyl-2-0-methy!-rac-glycero-3-phosphocholine, ET-I8-OCH 3 ), ilmofosine (BM 41.440), miltefosine (hexadecylphosphocholine, HePC), perifosine (D-21266), erucylphosphocholine (ErPC), or erufosine (ErPC3,

erucylphosphohomocholine; or an indole-3 -carbinal analogue such as indole-3 -earbinol, 3- chloroacetylindole, diindolylrnethane, diethyl 6-methoxy-5,7-dihydroi dolo [2,3~/>]earbazole~ 2,10-dicarboxylate (SR13668), or OSU-A9; a sulfonamide derivative (e.g, PH-316, PHT-427); a thiourea derivative such as PIT-1, PIT-2, DM -PIT- 1 , or N~[(l -methyl- Ii/-pyrazol-4~ yl)carbonyl]-N'-(3-bromophenyl)-thiourea; a purine derivative such as triciribine (TCN, NSC 154020), triciribine mono-phosphate active analogue (TCN-P), 4-amino-pyrido[2,3- i jpyrimidine derivative API-1, 3~phenyl~3i7-imidaz.o[4,5~/)]pyridine derivatives, or ARQ 092; BAY 1125976, 3-methyl-xanthine, quinoline-4-carboxamide and 2-[4-(cyclohexa-l,3-dien-l- yl)-17/-pyrazol-3-yl]phenol, 3-oxo-tirucallic acid, 3a- and 3p~acetoxy-tirucallic acids, or acetoxv-tirucallic acid. In some embodiments, the AKT inhibitor is an irreversible inhibitor, such as a natural product, an antibiotic such as lactoquinomycin, frenolicin B, kalafungin, medermycin, Boc-Phe-vinyl ketone, 4-hydroxynonenal (4-HNE), 1,6-naphthyridinone derivatives, or an i midazo- 1 ,2-pyridine derivative.

[0117] in some embodiments, the mTOR inhibitor is a rapamycin-like nutraceutical agent. A rapamycin-like nutraceutical agent is an agent, or precursor or derivative thereof, which exhibits anti -mTOR pathway effects. Exemplary' rapamycin-like nutraceutical agents include, but are not limited to, agents such as steroids, resveratrol, pterostilbene, aspirin, polyunsaturated fatty acids, curcumin, R-lipoic acid, caffeine, fisetin, apigenin, alcohol, andrographis, pomegranate, Reishi, milk thistle, oieanolic acid, anthocyanins, Astragalus, Rhodiola, Camosine, Plumbagin and Chrysophanic acid

[0118] In some embodiments, the method comprises administration of an effective amount of one or more mTOR inhibiting agents in cycles to achieve a high level of mTOR inhibition. In some embodiments, the one or more mTOR inhibiting agents can inhibit mTOR expression or activity by at least about any one of 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or more. For example, rapamycin or a rapamycin-like nutraceutical agent and rapalogs are known to be potent mTOR inhibitors that can achieve a high level of mTOR inhibition, such as niTORCl inhibition. The percentage of inhibition can be measured by methods such as, but not limited to, the quantity of proteins ofmTORCl suppression from the pS6K/pS6 axis, and also in-vitro cellular re-proliferative potential, senescent cell hypertrophy, morphology, Oil Red O staining and yeast cell chronological life span (CLS) measurement. In some embodiments, the one or more mTOR inhibiting agents are administered weekly or bi-weekly. In some embodiments, each cycle of administration comprises seven weeks, and the one or more mTOR inhibiting agents are administered for six weeks and off in week 7. In some embodiments, each cycle of

administration comprises eight weeks, and the one or more mTOR inhibiting agents are administered for six weeks and off in week 7 and week 8.

[0119] In some embodiments, the cyclic regimen of the one or more mTOR inhibiting agents is combined with administration of an effective amount of one or more mTOR inhibiting agents to maintain a modest baseline threshold of mTOR inhibition. In some embodiments, the one or more mTOR inhibiting agents can inhibit mTOR activity by no more than about any one of 40%, 30%, 20%, 10%, 5% or less. For example, direct and indirect AMPK activators are suitable for maintaining a modest baseline threshold of mTOR inhibition. Many known methods and kits can be used to assess mTOR activity. As non-limiting examples, ELISA and western blots can be used to assess the kinase activity of immunoprecipitated mTOR from cell lysates on its substrates

[0120] Cyclic administration can reduce undesirable toxicities of some highly potent mTOR inhibiting agents that lead to a constantly high level of mTOR inhibition. Such toxicities, including hematological toxicity and mucosal ulcers, etc , have been observed when mTOR inhibitors are used as immune suppressant in kidney transplant and cancer treatments. These toxicities are undesirable for long-term anti-aging interventions. In some embodiments, w'herein the mTOR inhibitor is administered locally, the toxicity effects may be localized, and cyclic administration of the mTOR inhibitor may not be necessary.

[0121] In some embodiments, the effective amount of the mTOR inhibitor is chosen to reduce undesirable side effects and toxicity of the mTOR inhibitor. In some embodiments, wherein the mTOR inhibitor comprises a rapamycin and/or a rapalog, and wherein the mTOR inhibitor is administered systemically, the effective amount of rapamycin or the rapalog is sufficient to maintain a systemic plasma or serum trough level of no more than about 8 ng/ml of rapamycin or equivalent amount of the rapalog. In some embodiments, the dose and dosing frequency of rapamycin and/or the rapalog are chosen to maintain a systemic plasma or serum trough level of no more than about 8ng/mi of rapamycin or equivalent amount of the rapalog. In some embodiments, wherein rapamycin is administered systemically, the dose of rapamycin is about any one of 0.5 mg to 8 mg daily, 1 mg to 15 mg once weekly, or about 2 mg to 40 mg once every 2 or 3 weeks. The trough level of rapamycin or the rapalog in plasma or serum samples can be monitored periodically to guide selection of an appropriate dosage.

[0122] In some embodiments, wherein the mTGR inhibitor comprises a rapamycin and/or a rapalog, and wherein the mTOR inhibitor is administered locally, the effective amount of rapamycin or the rapalog is sufficient to maintain a systemic plasma or serum trough level of no more than about 2 ng/ml of rapamycin or equivalent amount of the rapalog. In some

embodiments, wherein the mTOR inhibitor comprises a rapamycin and/or a rapalog, and wherein the mTOR inhibitor is administered via a combination of local and systemic routes, the effective amount of rapamycin of the rapalog is sufficient to maintain a systemic plasma or serum trough level of no more than about 10 ng/ml.

[0123] In some embodiments, the mTOR inhibitor also functions as an insulin sensitizer.

C. Senolytic p53/pRb activator

[0124] The methods described herein comprise administering an effective amount of a senolytic p53/pRb activator, which may comprise one or more agents that promotes p53 and/or pRb activity. In some embodiments, the senolytic p53/pRb activator comprises an agent

(referred herein as p53 activator”) that activates p53, and/or stabilizes p53. In some

embodiments, the senolytic p53/pRb activator comprises an agent (referred herein as“pRb activator) that activates pRb, stabilizes pRb, and/or dephosphoryiates pRb. The pRb activators can also activate, and/or stabilize p53. In some embodiments, the senolytic p53/pRb activator comprises both a p53 activator and a pRb activator. There is extensive cross talk between downstream effector molecules of the p53 and pRb pathways. In some embodiments, the senolytic p53/pRb activator is a single agent that leads to increased p53/pRb activities. In some embodiments, the senolytic p53/pRb activator increases p53 activity by at least about any one of 20%, 50%, 75%, 100%, 150%, 200% or more. In some embodiments, the senolytic p53/pRb activator increases pRb activity by at least about any one of 20%, 50%, 75%, 100%, 150%, 200% or more. Many known methods and kits can be used to assess p53 and pRb activity, including, but not limited to, ELISA and western blots that can be used to assess the levels of p53 and pRb. Also, RT-PCR can be used to assess the levels of mRNA transcripts of genes under the control of p53 and pRb to determine the activity of p53 and pRb.

[0125] In some embodiments, the senolytic p53/pKb activator comprises an agent that activates p53. In some embodiments, the senolytic p53/pRb activator comprises an agent that inhibits or suppresses binding of p53 to MDM2 or MDMX. Suitable inhibitors of p53-MDM2 binding or p53-MDMX binding include, but are not limited to, nutlins such as RG7112 (i.e., RO5045337), RO5503781 ; benzodiazedinediones such as TDP665759; spiro-oxindoles such as MI-219 and SAR405838; imidazothiazole such as DS-3032b; dihydroisoquinolinone such as CGM-097; HDM201; piperidines such as MK4828; piperidinone such as AMG232; pyrroidine such as RG7388; peptide MDM2/X inhibitors such as ALRN-6924, ALRNRITA; and other small molecule inhibitors such as JNJ-26854165 (i.e., serdemetan), SJ-172550, RO-2443/RO- 5693, and XI -01 1.

[0126] In some embodiments, the senolytic p53/pRb activator comprises an agent that inhibits a sirtuin, such as SirTl and/or SirT2. Suitable sirtuin inhibitors include, but are not limited to, Tenovin 1 and 6

[0127] In some embodiments, the senolytic p53/pRb activator comprises a COX inhibitor. Without being bound by any theory or hypothesis, COX inhibitors can promote or stabilize p53 transcription by down-regulating cycloxygenase-2 (COX-2), retrieve p53 from COX-2 association, and/or activate p53 via ataxia telangiectasia mutated-/p38 mitogen-activated protein. Upon genotoxic stress, COX-2 and p53 accumulate in the nucleus, where they physically interact with one another. An amino-terminal region (amino acids 1—126) of COX-2 interacts with the DNA-binding domain of p53. The p53-interacting region is critical for CQX-2- mediated inhibition of p53, including DNA binding, transcriptional activity, and p53- and genotoxic stress-induced apoptosis. In addition, an active site mutant of COX-2 (S5 I6Q) as well as wild-type COX-2 potently inhibits p53 transcriptional activity and genotoxic stress-induced apoptosis. These results suggest that COX-2 can inhibit p53 function through a catalytic activity- independent mechanism, and that COX-2 inhibits p53 function through physical interaction with p53 in the nucleus.

[0128] In some embodiments, the senolytic p53/pRb activator comprises a nonselective COX inhibitor. Exemplary non-selective COX inhibitors, include, but are not limited to, acetates such as diclofenac, indomethacin, and sulindac; fena ales such as mefenamic acid; oxicams such as piroxicam; propionates such as ibuprofen, ketoprofen, naproxen; pyrazolones such as

phenylbutazone; salicylates such as aspirin, and diunisal. In some embodiments, the senolytic p53/pRb activator comprises a selective COX inhibitor. Exemplary selective COX inhibitors include, but are not limited to, meloxicam, nimesu!id, and coxibs such as first generation celecoxib, second generation celecoxib, etorcoxib and valdecoxib.

[0129] In some embodiments, the senolytic p53/pRb activator comprises a COX inhibitor having a half-life less than about 6 hours, e.g., aspirin, diclofenac, ibuprofen, indo ethacin, or ketoprofen. In some embodiments, the senolytic p53/pRb activator comprises a COX inhibitor having a half-life more than about 10 hours, e.g., diunisal, naproxen, phenylbutazone, piroxicam, or sulindac.

[0130] In some embodiments, the senolytic p53/pRb activator comprises an agent that activates pRb. Exemplar}' agents that can activate pRb include, but are not limited to, agents that dephosphorylate pRb; pan CDK inhibitors such as flavopiridol, olomoucine II, purvalanol A, SNS-032 (Le., BMS387032), dinaciclib, MK-7965, SCH727965, AT7519 (Astex), R547, AZD5438, and AG024322; and specific CDK4/6 inhibitors, such as ribociclib, palbociclib (i.e., PD-0332991), abemaciclib (i.e., LY2835219, VERZENIO™), and trilaciclib (i.e., G1 T28).

[0131] The effective amount of the senolytic p53/pRb activator can be chosen based on effective dosages of the senolytic p53/pRb activator established in the art. The effective dosages of many COX 2 inhibitors for systemic and/or local administration have been established. For example, the recommended safe daily oral dose of ibuprofen is no more than about 1200 mg.

[0132] Senolytic (from the words "senescence" and "lytic" - destroying) molecules can selectively induce death of senescent cells. Without being bound by any theory or hypothesis, senescence is a potential tumor suppressive mechanism and possible factor that accelerates the aging process. Senolytic agents or senolytic regimen are designed to delay, prevent, alleviate, ameliorate, or reverse age-related diseases. Certain anti -cancer agents in low doses can decelerate aging and age-related diseases as these agents generally remove older cells.

Targeting cancer prevention pathways with anti -cancer agents may confer longevity effects by offering protection from metabolic pathologies during aging, independently of effects on cancer.

[0133] Without wishing to be bound by any theory or hypothesis, aging at cellular level leads to cell senescence, which contributes profoundly to whole-body aging. Elimination of senescent cells has potential to increase human life expectancy. Senolytic drugs or regimens are agents that can kill senescent cells with minimal harm to normal cells. Certain senolytic agents promote apoptosis. Apoptosis is cel! suicide, and apoptosis of senescent cells is beneficial to the full organism. Removal of senescent cells is an appealing approach for anti-aging therapy. Senolytic strategy finds support from empirical data in mammals, parabiosis experiments, and the theory that circulating chemical signals form the basis of an epigenetic clock. Some of these circulating molecules are known to come from senescent cells. Another theory is that aging accelerates exponentially with age, as though it rvere driven by a positive feedback loop. For

example, senescent cells may secrete cytokines that make more senescent cells, thereby forming a positive feedback loop that accelerates aging. In addition, short telomeres initiate

senescence. At any given time, there is a bell-shaped curve of telomere length among the body’s cells. The tail of the telomere distribution contains a few cells that are driven to senescence by having very short telomeres. As organisms age, more and more of these senescent cells accumulate, which may in turn support the positive feedback loop that accelerates aging.

[01341 It is difficult to formulate a senolytic regimen that works without harming normal cells. For example, FOXO-DRI is a senolytic agent that dissociates FQXQ4. FOXO is a master transcription factor associated with aging and development. It is the mammalian homolog of the pivotal life extension protein first identified in worms as DAF16 in the 1990s. FOX04 activation in a cel! can block apoptosis. p53 is the most common trigger of apoptosis, the first protein biochemists usually think of in connection with apoptosis. p53 has multiple functions in the cell nucleus, but as a trigger for apoptosis, it functions through the mitochondria. FOX04 binds to p53 and blocks its induction of apoptosis. The efficacy of a senolytic agent can be measured by its ability to kill senescent cells without doing harm to normal cells. The index called SI 50 (SI for“selectivity index - 50%”) is defined by analogy to LD 50 = the“lethal dose” of a toxin, the dose at which half of all cells die. SFo is defined as the ratio of LD JO ’S for normal and senescent cells. It is the concentration of the agent at which half the normal cells die, divided by the concentration at which half the senescent cells die. The FOX04 blocking agent FOXO-DRI has a SI50 about 12, which can leave a lot of normal cells live after usage, but is not high enough to support a useful therapy. After a standard dose of FOXO-DRI is injected in humans, the cellular concentrations vary from person to person and from tissue to tissue. If the 8I 50 is low in vital tissues and organs, then toxic damage similar to chemotherapy can occur, preventing further development of such an agent.

[0135] FOXO-DRI is the newest senolytic agent, and it has the best ratio yet for killing senescent ceils while avoiding collateral damage to healthy cells. It cannot be taken orally and must be injected. However, because of expected toxicities, FOXO-DRI has been developed mainly as an anti-cancer agent rather than an anti-aging agent. Dasatinib and Quercetin (not via the p53 pathway) were also developed as an early proof-of-principle senolytic regimen, and both drugs were FDA approved. However, the combination was found to be too toxic to be effective. Therefore, prior to this invention, no senolytic regimen via p53~depenent or p53-independent pathway has been successfully applied for anti-aging purposes.

D. NAD+ supplement

[0136] The methods described herein comprise administering an effective amount of a NAD+ supplement, which may comprise one or more agents that increases the level or maintains a high level of NAD+. In some embodiments, the NAD · level in the body (e.g, blood, tissue or cellular level) is maintained at about 500 uM. In some embodiments, the NAD+ level in the body is maintained at least about any one of 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, 600 mM, 700 mM, 800 mM, 900 mM, 1 mM, or higher.

[0137] Without being bound by any theory or hypothesis, an adequate rate of NAD+

generation and maintenance of a high level of NAD+ can increase the body’s N AD availability, which provides adequate NAD+ to promote activation of Sirtuins (Sirtuins 1-7) even when P ARP i is activated. The NAD+ concentration available before PART 1 activation can be depleted by more than 80% upon PARP1 activation. In some embodiments, the NAD+ level is maintained at the level when PARP1 is not activated in order to activate Sirtuins. Activation of PARP1 and Sirtuin can be assessed using known methods in the art, such as ELISA, and

Western blotting. The NAD+ level in ceils, tissues, and blood samples can be determined using enzymatic or cell-based colorimetric or fluorescence assays.

[0138] In animal models, a single NAD+ supplement can reverse dyslipidemia and diseases associated thereof. In some embodiments, a single agent that increases NAD+ supply is not sufficient to initiate and achieve cellular repair and ultimate prolonging survival. For example, chronic inflammation from various causes together with aging, may override the benefits of a single agent supplement approach, which may be insufficient to sustain a high level of NAD+ for rate limiting enzymatic pathways. The human body responds to aging via complicated positive and negative feedback processes, such as genetic and epigenetic transcriptions, pre- and post- translational modifications, stimulatory ' and inhibitory ' small molecules, peptides, and proteins, and the presence or absence of pre-existing metabolic conditions. Most of these processes are either continuous or occurring in rapid self-reactivation downward spiraling cycles. Simply supplying NAD+ using methods and routes known in the art may be insufficient to provide sustainable and effective NAD+ levels to override this long established aging process.

[0139] In some embodiments, the NAD+ supplement comprises niacin, i.e. nicotinic acid (“NA”). NA is from the de novo crossover pathway. A suitable dosage of NA is at least

250mg/day for a 60 Kg adult. In some embodiments, the NAD+ supplement comprises one or more of nicotinamide riboside (“NR”), nicotinamide mononucleotide (“NMN”), and

nicotinamide (NAM). NR, NMN and NAM are agents of the NAD salvage pathway. A suitable dosage of NR, NMN, and NAM can be chosen based on a moles/L equivalent to niacin, such as about 500mg/day for NR, about 750mg/day for NMN, and about 250mg for NAM for a 60Kg adult.

[0140] In some embodiments, the NAD+ supplement comprises a combination of two or more (e.g., 2, 3, 4, or more) agents from different NAD+ generation pathways, which have different pharmacokinetics, pharmacodynamics, and rates of rise of NAD+ in different tissues. In some embodiments, the combination of agents maintains a high level of NAD+ in ceils or tissues of the body, thereby mitigating the aging process and its associated diseases. In some embodiments, the NAD+ supplement comprises a combination of agents from both NAD de novo crossover pathway and the NAD salvage pathway.

[0141] NA and NR are among the most effective and bioavailab!e supplements to increase NAD+ and ADPR. See, Trammel et a!. Nature Communications 7: 12948 (2016). NR can be converted to NAAD through a yet unknown pathway. In some embodiments, the NAD+ supplement comprises a combination of NA and NR, which produce a high and constant level of NAD+. The methods described herein can maintain a NAD+ level over 500mM throughput cycles of NAD+ consumption, such as PARP1 activation.

[0142] In some embodiments, the NAD+ supplement consists of one or more agents of the de novo crossover pathway of NAD+ generation. In some embodiments, the NAD+ supplement consists of one or more agents of the salvage pathway of NAD+ generation. In some

embodiments, the NAD+ supplement comprises agents from both the de novo crossover pathway and the salvage pathway of NAD+ generation. In some embodiments, the NAD+ supplement consists of nicotinic acid (NA). In some embodiments, the NAD+ supplement comprises: (a) NA, and (b) one or more agents selected from NR, NMN and NAM. In some embodiments, the NAD+ supplement consists of NA and NR. In some embodiments, the NAD+ supplement consists of NA and NMN. In some embodiments, the NAD+ supplement consists of NA and NAM. In some embodiments, the NAD+ supplement consists of NA, NR and NMN. In some embodiments, the NAD+ supplement consists of NA, NR and NAN. In some embodiments, the NAD+ supplement consists of NA, NMN and NAN. In some embodiments, the NAD+ supplement consists of NA, NR, NMN and NMN. In some embodiments, the NAD+ supplement does not comprise NA. In some embodiments, the NAD+ supplement consists of one or more agents selected from NR, NMN and NAM.

[0143] In some embodiments, the NAD+ supplement is administered systemically, such as orally. The effective amount of the NAD+ supplement for systemic administration is sufficient to maintain NAD+ at a level of about 200 mM to about 500 iiM. In some embodiments, the effective amount of the NAD+ supplement is sufficient to restore NAD+ to at least about any one of 50%, 60%, 70%, 80%, 90%, 100% or more of baseline level (e.g., 200 mM to about 500 mM) post P ARP 1 activation. Suitable dosages for oral administration include, for example, at least about 250 mg/day of Nicotinic acid (NA) in combination with: any one of (a) at least about 5Q0mg/day of nicotinamide riboside (NR), (b) at least about 750mg/day of nicotinamide mononucleotide (NMN), and (c) at least about 250mg/day of nicotinamide (NAM).

[0144] In some embodiments, the NAD+ supplement is administered locally, such as topically. The effective amount of the NAD+ supplement for local administration is sufficient to maintain NAD+ at a level of more than about 1 mM. In some embodiments, the effective amount of the NAD · supplement is sufficient to restore NAD+ level to more than about 200% of baseline level (e.g, to increase the NAD+ level from less than about 200 mM to more than about ImM) post P ARP 1/2 and CD38/157 activation. Suitable dosages of each component of the NAD+

supplement can be chosen based on an estimated distribution volume, pharmacokinetics, and penetration of the component to achieve a minimum of about 100 mg of NAD+ per 100 ml of body tissue targeted.

[0145] For example, an estimated 38.46 mg/100 ml (the first calculation) of topical nicotinic acid (NA) solution is needed assuming 100% penetration, distribution and absorption. Drug transport in the skin is a process involving several steps, including dissolution and release of drug from the formulation, partitioning of the drug into the stratum corneum, diffusion of the drug across the stratum corneum (e.g, by intercellular lipids), partitioning of the drug from the stratum corneum into viable epidermis layers, diffusion across the viable epidermis layers into the dermis, and absorption of the drug by capillary' vessels, which achieves systemic circulation (Kalia, Guy, 2001). Taking the above variable reasons and using models of transdermal FDA- approved and similar molecular weight chemicals (compared to nicotinic acid and nicotinamide riboside) such as nitroglycerine a second calculation yields a total delivered dose of about 40mg. This presumption has a 20ug/ cnri/hr penetration, yielding a final plasma concentration of 1 nM with a half-life of 15 minutes, and assuming total delivery of NAD+ supplements is no more than about 18% of the topical formula. The first calculation of 38.46 mg/100 mi is very' close to the value of 40mg in the second calculation. In some embodiments, the NAD+ supplement is formulated for local administration, and the NAD+ supplement comprises a minimum total daily dose of no less than about 10ml of 0.3% of Nicotinic acid (NA) by itself or in combination with any one or a combination of the three agents of the salvage pathway, a minimum total daily dose of no less than 10 ml of 0.6% of nicotinamide riboside (NR), a minimum total daily dose of no less than 10 ml of 0.9% of nicotinamide mononucleotide (NMN), or a minimum total daily dose of no less than 10ml of 0.3% of nicotinamide (NAM).

E. Administration

[0146] Each of the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, seno!ylic p53/pRb activator, and NAD+ supplement described herein can be administered using a suitable route of administration, including systemic and local admini stration. In some embodiments, one or more of the insulin sensiti zer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and NAD+ supplement is administered locally. In some embodiments, one or more of the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and NAD+ supplement is administered systemically. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, or NAD+ supplement, which comprises two or more agents, is administered by a combination of local and systemic routes, e.g., one agent is administered locally and another agent is administered systemicaliy. Exemplary routes of administration include, but are not limited to, intravenous, intra-arterial, intraperitoneal, intrapulmonary, intravesicular,

intramuscular, intra-tracheal, subcutaneous, intraocular, intrathecal, intradermal, transdermal, rectal, vaginal, intra-cavity, topical and oral.

[0147] In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement are administered systemicaliy. Suitable systemic routes include, but are not limited to, intravenous, intramuscular, subcutaneous administration. In some embodiments, the insulin sensitizer, mTQR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, seno!ytie p53/pRb activator, and/or NAD+ supplement is via gastrointestinal route, such as orally. In some embodiments, gastrointestinal administration is preferred for long term treatment.

[0148] In some embodiments, local including regional administration is used. Compared to systemic administration, local administration can achieve different phar acokinetics and pharmacodynamics for different components in combination regimen (i.e., the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and NAD+ supplement). For example, metabolism and

modifications by the gastrointestinal route and the first pass effects through the liver, coupled with systemic toxicides for higher doses can pose challenging hurdles for long-term systemic administration of the combination regimen. The advantages of local administration of certain agents described herein, e.g, NAD supplements, pterostilbene or resveratrol etc., is numerous. For example, Sirtuins 1-7 cannot be activated by allosteric molecules such as resveratrol and NAD+ unless certain higher than orally achievable level is reached.

[0149] Local administration can be achieved using any suitable device, media, methodology, and route known in the art. Local administration can achieve minimal or reduced toxicity than systemic delivery of the same agent. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is formulated in a topical media such as a cream, gel, lotion, solution, serum etc. for local administration, e.g., through skin or a mucous membrane. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pKb activator, and/or NAD+ supplement is formulated with one or more agents that facilitate local delivery', e.g., enhance penetration through skin or a mucous membrane. Suitable agents that facilitate local delivery include, but are not limited to, liposomes, dendrimers and micro-emulsions or other chemical enhancers. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is locally administered via a physical method, such as heat thermal abrasion, passive, iontophoresis, electroporation, cavitational ultrasound, or

microdermabrasion/microneedles. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is locally administered via an injection route, for example, intraderma! , subcutaneous, intramuscular or by injection into an organ or tissue. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is locally administered via a vascular delivery route, such as intra-arterial, via an arterial occlusion device, and/or via catheterization. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is locally administered via an intra-cavity route, e.g., by instillation into one or more specific body cavities. Suitable intra-cavity routes of administration include, but are not limited to, intravesical, intra-peritoneal, intra-pleural, intra-crania!, intra nasal, retrograde and intra-ocular or topical ocular procedures. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is locally administered via a device implanted into a tissue or organ, such as a drug eluting stent, an intradermal pouch, a transderma! patch or membrane, a matrix, a gel and a reservoir (e.g., CT, MRI, or X-ray guided reservoir), etc.. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is locally administered via a suppository' preparation for rectal or vaginal application.

[0150] Suitable routes of administration and formulations for each agent in the combination regimen are chosen depending on the nature of the aging-related disease or condition being treated or prevented. Section“F. Indications” below further provide exemplary administration routes and methods that are suitable for particular aging-related diseases and conditions.

[0151] Each of the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and N AD supplement may be administered using any suitable schedule. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is administered daily or twice daily. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is administered is administered at least about any one of lx, 2x, 3x, 4x, 5x, 6x, or 7x (i.e., daily) a week. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is administered weekly. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is administered weekly without break; weekly, two out of three weeks; weekly three out of four weeks; weekly six out of seven weeks, once every two weeks; once every 3 weeks; once every 4 weeks; once every 6 weeks; once every' 8 weeks, monthly, or every two to 12 months ln some embodiments, the intervals between each administration are less than about any one of 6 months, 3 months, 1 and ½ month (6 weeks), 1 month, 20 days, 15, days, 12 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the intervals between each administration are more than about any one of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months. In some embodiments, there is no break in the dosing schedule. In some embodiments, the interval between each administration is no more than about a week. In some embodiments, the insulin sensitizer, mTQR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and NAD+ supplement are administered with the same dosing schedule. In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRh activator, and NAD+ supplement are administered with different dosing schedules.

[0152] The methods described herein may involve a single treatment, or repeated treatments.

In some embodiments, the combination regimen is administered for any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50 or more cycles. Each cycle of administration may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more weeks. In some embodiments, administration of the combination regimen is repeated once per week, once 2 weeks, once 3 weeks, once 4 weeks, once per month, once per 2 months, once per 3 months, once per 4 months, once per 5 months, once per 6 months, once per 7 months, once per 8 months, once per 9 months, or once per year. In some embodiments, the interval between each cycle of administration is about any one of 1 week to 2 weeks, 2 weeks to 1 month, 2 weeks to 2 months, I month to 2 months, 1 month to 3 months, 3 months to 6 months, or 6 months to a year. In some embodiments, the combination regimen is administered over a period of time, such as at least about any one of 1 month, 3 months, 6 months, 1 year, 2 years, 3 years,

5 years, 10 years, or more. F. Indications

[0153] The methods and compositions described herein are useful for treating or preventing a variety of aging-related diseases and conditions in any individual in need thereof. Non-limiting examples of age-related diseases include cardiovascular diseases (e.g., stroke, atherosclerosis, hypertension, heart failure), osteoporosis, insulin-resistance and type II diabetes (e.g., diabetic retinopathy, neuropathy), CNS diseases (e.g., Alzheimer's disease, Parkinson's disease), ocular diseases (e.g., age-related macular degeneration, cataracts, retinopathy), skeletal muscular diseases, skin diseases and conditions (e.g. seborrheic keratosis, actinic keratosis, hair loss and alopecia, photo-aged skin, wrinkles, and skin spots), auto-immune diseases (e.g., systemic lupus erythematosus, psoriasis, eczema, arthritis), and primary or metastatic cancer lesions (such as skin cancer and benign prostatic hyperplasia, e.g., for prevention or as adjuvant therapy).

[0154] In some embodiments, the insulin sensitizer, mTQR inhibitor, agent having dual functions as an insulin sensitizer and an mTQR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement are administered systemically (e.g., orally). In some embodiments, the aging- related disease or condition is a chronic disease or condition due to aging. In some embodiments, the aging-related disease or condition is diabetes, cardiovascular diseases (e.g., stroke, heart failure and atherosclerosis), neurodegenerative diseases (e.g, peripheral or CNS

neurodegenerative disease, such as Alzheimer’s disease), metabolic syndrome (e.g., fatty liver), osteoarthritis, or age-related macular degeneration (AMD). In some embodiments, the individual has a combination of any of the above aging-related diseases or conditions. In some

embodiments, the insulin sensitizer comprises metformin or an insulin sensitizing nutraceutical agent. In some embodiments, the insulin sensitizer comprises TZD. In some embodiments, the TZD is administered at a dosage lower than the dosage for treating bladder cancer and cardiac failure. In some embodiments, the insulin sensitizer comprises TZD and metformin or an insulin sensitizing nutraceutical agent. In some embodiments, the mTQR inhibitor is a rapalog, such as everolimus. In some embodiments, the mTOR inhibitor comprises a pan-mTGR inhibitor (e.g., torin 2). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor, a CDK4/6 inhibitor, a pan-CD K inhibitor, and combinations thereof. In some embodiments, the senolytic p53/pKb activator is nutlin. In some embodiments, the senolytic p53/pRb activator is a COX2 inhibitor, such as voltaren, ibuprofen or aspirin. In some embodiments, the senolytic p53/pRb activator is a CDK4/6 inhibitor and/or a pan CDK inhibitor. In some embodiments, the NAD+ supplement comprises: (a) NA, and (b) one or more of NR, NMN and NAM.

[0155] In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD · supplement are administered locally (e.g, topically). In some embodiments, the aging- related disease or condition is a skin disease, such as aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss and alopecia, photo-aged skin, wrinkles, skin spots, skin cancer (e.g., for prevention or as adjuvant therapy), psoriasis, or eczema. In some embodiments, the method is for cosmetic applications, such as for treating or preventing wrinkles. In some embodiments, the aging-related disease or condition is a disease or condition of tendons, ligaments, or muscle groups, such as muscle, tendon and joint pain, inflammation or strain, including all forms of arthritis such as osteoarthritis and rheumatoid arthritis. In some embodiments, the individual has a combination of any of the above aging-related diseases or conditions. In some embodiments, the age-related disease or condition is metabolic disease, diabetes or pre-diabetes, non-alcoholic fatty liver disease, non-alcoholic steatotie hepatitis, fat reduction or“sculpture” procedures such as the surgical removal of fat using liposuction and/or the application of cold, heat, laser, ultrasound, radiofrequency ablation.

[0156] In some embodiments, the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement are administered locally, such as by instillation or implantation of a local delivery device, matrix or gel in an aging sensitive tissue or organ or a focal lesion thereof. In some embodiments, the aging-related disease is chronic cystitis, and wherein the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement are administered via an intravesical gel. In some embodiments, the aging-related disease or condition is a disease in the central nervous system (e.g., Amyotrophic lateral sclerosis or Parkinson’s disease), and wherein the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement are administered intracranially or intrathecally. In some embodiments, the aging-related disease or condition is a cardiovascular disease (e.g. myocardial infarction, heart failure, or coronary heart disease), wherein the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement is administered via a drug-eluting stent. In some embodiments, the aging-related disease or condition is an intra-ocular disease (e.g., AMD), and wherein the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pKb activator, and/or NAD+ supplement are administered via an intra-ocular device or as an eye drop solution. In some embodiments, the aging-related disease or condition is diabetes or liver failure, and wherein the insulin sensitizer, mTOR inhibitor, agent having dual functions as an insulin sensitizer and an mTOR inhibitor, senolytic p53/pRb activator, and/or NAD+ supplement are administered via CT, MRI, or X-ray guided reservoir or device insertion.

[0157] In some embodiment, wherein the aging-related disease or condition is cancer, the method is for prevention or used in adjuvant setting.“Adjuvant setting” refers to a clinical setting in which an individual has had a history of cancer, and generally been responsive to therapy, which includes, but is not limited to, surgery, radiotherapy, and chemotherapy.

Treatment or administration in the“adjuvant setting” refers to a subsequent mode of treatment. G. Kits and pharmaceutical compositions

[0158] The present application also provides compositions (such as pharmaceutical compositions), kits and articles of manufacture for carrying out any one of the methods described herein.

[0159] In some embodiments, there is provided a pharmaceutical composition comprising: (i) an insulin sensitizer, (ii) an mTOR inhibitor; (iii) a senolytic p53/pRb activator, and (iv) an NAD+ supplement; and a pharmaceutically acceptable excipient hi some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan- mTOR inhibitor (e.g, torin I, torin 2, PP242 or INK128). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD-ί- supplement comprises NA, NR, NMN, and/or NAM.

[0160] In some embodiments, there is provided a pharmaceutical composition comprising: (i) agent having dual functions as an insulin sensitizer and an mTOR inhibitor, (ii) a senolytic p53/pRb activator; and (iii) an NAD+ supplement; and a pharmaceutically acceptable excipient. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin in some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM.

[0161] In some embodiments, there is provided an implantable device for treating or preventing an aging-related disease or condition, comprising: (i) an insulin sensitizer; (ii) an rnTOR inhibitor; (iii) a senolytic p53/pRb activator, and (iv) an NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., piog!itazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutieal agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutieal agent, and/or a rapalog (e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan-mTOR inhibitor (e.g., torin 1, torin 2, PP242 or INK 128). In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof.

In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiments, the implantable device is selected from the group consisting of an intravesical or other urogenital tract gel, an intra-cranial or intra-thecal device, intra-ocular device, a drug eluting stent, and CT, MRI or X-ray guided and inserted devices.

[0162] in some embodiments, there is provided an implantable device for treating or preventing an aging-related disease or condition, comprising: (i) agent having dual functions as an insulin sensitizer and an mTOR inhibitor, (ii) a senolytic p53/pRb activator; and (iii) an NAD+ supplement; and a pharmaceutically acceptable excipient. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin. In some embodiments, the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM.

[0163] For use in prophylaxis and/or therapy of aging-related diseases via the local and/or systemic routes, compositions described herein can be administered in a conventional dosage form prepared by mixing with a standard pharmaceutically acceptable carrier according to known techniques. Some examples of pharmaceutically acceptable carriers can be found in: Remington: The Science and Practice of Pharmacy (2005) 21 st Edition, Philadelphia, Pa. Lippincott Williams & Wilkins. The compositions may be provided as pharmaceutical preparations, examples of which include, but are not limited to mixtures, solutions, creams, liniments, eye drops, and nanoparticle compositions.

[0164] In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is formulated in a pharmaceutical composition comprising a

pharmaceutically acceptable excipient. In some embodiment, the insulin sensitizer and the

TOR inhibitor, or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor; the senolytic p53/pRb activator and the NAD+ supplement are formulated in a single pharmaceutical composition. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions. In some embodiments, the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and the NAD+ supplement are each formulated in a different matrix for immediate or extended release. In some embodiments, the pharmaceutically acceptable excipient is a serum, gel, buffer solution, cream, lotion, liniment, ointment or combinations thereof.

[0165] In some embodiments, the insulin sensitizer or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor comprises metformin. In some embodiments, the insulin sensitizer is in a pharmaceutical composition comprising at least about 0.5% (w/v), at least about 0.7% (w/v), or 0.5-10% (w/v) metformin. In some embodiments, the insulin sensitizer comprises pioglilazone. in some embodiments, the insulin sensitizer is in a

pharmaceutical composition comprising at least about 0.03% (w/v) pioglitazone. In some embodiments, the mTOR inhibitor comprises rapamycin In some embodiments, the mTOR inhibitor is in a pharmaceutical composition comprising at least about 0.01% (w/v) rapamycin.

In some embodiments, the senolytic p53/pRb activator comprises aspirin. In some embodiments, the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 1% (w/v) aspirin. In some embodiments, the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0.5% (w/v) or about 0.5-4% (w/v) diclofenac. In some embodiments, the senolytic p53/pRb activator comprises nutiin-3. In some embodiments, the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0 3% (w/v) nutiin-3. In some embodiments, the NAD+ supplement is in a pharmaceutical composition comprising; (a) at least about 0.5% (w/v) Nicotinic Acid, and (b) at least about 1% (w/v) Nicotinamide Riboside (NR), at least about 1.5% (w/v) Nicotinamide Mononucleotide (NMN), and/or at least about 0.5% (w/v) Nicotinamide (NAM). In some embodiments, the NAD+ supplement is in a pharmaceutical composition comprising at least 0.5 % (w/v) or about 0.5-6% (w/v) NR. In some embodiments, the NAD+ supplement is in a pharmaceutical composition comprising at least 0.5 % (w/v) or about 0 5-6% (w/v) NA. In some embodiments, the pharmaceutical composition further comprises vitamin D, vitamin E, vitamin A, resveratrol, pterosti!bene, L-carnitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Chayote ( Sechium edule) juice or extract. In some embodiments, the pharmaceutical composition comprises at least 0.5 % (w/v), such as about 0.5-3% (w/v) pterostilbene. In some embodiments, the

pharmaceutical composition further comprises about 1-10% (w/v) urea, about 1 -10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40% (w/v) PEG200, and about 5-40% (w/v) PEG400.

[0166] In some embodiments, there is provided a kit for treating or preventing an aging-related disease or condition, comprising: (i) an insulin sensitizer, (ii) an rnTOR inhibitor; (iii) a senoiytic p53/pRb activator; and (iv) an NAD+ supplement. In some embodiments, the insulin sensitizer comprises a TDZ (e.g., pioglitazone) and/or a biguanide (e.g., metformin). In some embodiments, the insulin sensitizer is an insulin sensitizing nutraceutical agent. In some embodiments, the mTOR inhibitor comprises rapamycin or a rapamycin-like nutraceutical agent, and/or a rapalog (e.g., everolimus). In some embodiments, the mTOR inhibitor comprises a pan- mTOR inhibitor (e.g, torin 1, torin 2, PP242 or INK128). In some embodiments, the senoiytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., ibuprofen), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD+ supplement comprises NA, NR, NMN, and/or NAM. In some embodiments, the kit further comprises an instruction for carrying out any one of the methods described above.

[0167] In some embodiments, there is provided a kit for treating or preventing an aging-related disease or condition, comprising: (i) agent having dual functions as an insulin sensitizer and an mTOR inhibitor, (ii) a senoiytic p53/pRb activator; and (iii) an NAD+ supplement; and a pharmaceutically acceptable excipient. In some embodiments, the agent having dual functions as an insulin-sensitizer and an mTOR inhibitor is metformin. In some embodiments, the senoiytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor (e.g., diclofenac), a CDK4/6 inhibitor, a pan-CDK inhibitor, and combinations thereof. In some embodiments, the NAD-- supplement comprises NA, NR, NMN, and/or NAM. In some embodiments, the kit further comprises an instruction for carrying out any one of the methods described above.

[0168] The kit may further comprise a description of selection of individuals suitable for treatment. For example, the kit may comprise a description of selection of individuals based on their HbAlC levels. Instructions supplied in the kits of the invention are typically written instructions on a label or package insert (e.g., a paper sheet included in the kit), but machine- readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable. The package insert contain instructions customarily included in commercial packages of therapeutic products that contain information about the indications, usage, dosage, administration, combination therapy, contraindications and/or warnings concerning the use of such therapeutic products.

[0169] The kits are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. Kits may optionally provide additional components such as buffers and interpretative information. The present application thus also provides articles of manufacture, which include vials (such as sealed vials), bottles, jars, flexible packaging, and the like.

[0170] Also provided are articles of manufactures comprising any one of the kits described herein. An“article of manufacture” is any manufacture (e.g , a package or container) or kit comprising at least one reagent, e.g., a medicament for treatment of a disease or disorder (e.g., aging-related disease or condition), or a probe for specifically detecting a biomarker (e.g., HbAlC) described herein. In certain embodiments, the manufacture or kit is promoted, distributed, or sold as a unit for performing the methods described herein.

[0171] Additionally, the arti cle of manufacture may further compri se a second container comprising a pharmaceuticaily-acceptabie buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

EXEMPLARY EMBODIMENTS

0172] Among the embodiments provided herein are:

1. A method of treating or preventing an aging-related disease or condition in an individual, comprising administering to the individual: (i) an effective amount of an insulin sensitizer, (ii) an effective amount of an mTOR inhibitor, (iii) an effective amount of a senolytic p53/pRb activator, and (iv) an effective amount of an NAD+ supplement.

The method of embodiment 1, wherein the insulin sensitizer comprises a

thiazo!idinedione and/or a biguanide.

The method of embodiment 2, wherein the thiazolidinedione is pioglitazone.

The method of embodiment 2 or 3, wherein the biguanide is metformin.

The method of any one of embodiments 1-4, wherein the mTOR inhibitor comprises rapamycin and/or a rapalog.

The method of embodiment 5, wherein the rapalog is selected from the group consisting of everolimus, tacrolimus, CC 1-779, ABT-578, AP-23675, AP-23573, AP-23841, 7-epi- rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl -rapamycin, 7-epi- thiom ethyl -rapamycin, 7-demethoxy-rapamycin, 32-demethoxy -rapamycin, 2-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desm ethyl -rapamycin, 42-0-(2- hydroxyjethyl rapamycin, and combinations thereof.

The method of any one of embodiments 1-6 wherein the mTOR inhibitor comprises a pan-mTOR inhibitor.

The method of embodiment 7, wherein the pan-mTOR inhibitor is selected from the group consisting of sapanisertib (INKI28), AZD2014, torin 1, torin 2, AZD8055, PP242, KU-006379, OSI-027, WAY-600, WAE-687, WYE-354, GSK1059615, and

combinations thereof.

A method of treating or preventing an aging-related disease or condition in an individual, comprising administering to the individual: (i) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor, (ii) an effective amount of a senolytic p53/pRb activator, and (iii) an effective amount of an NAD+ supplement. The method of embodiment 9, wherein the agent having dual functions as an insulin- sensitizer and an mTOR inhibitor is metformin.

The method of any one of embodiments 1-10, wherein the senolytic p53/pRb activator is selected from a nutlin or analog thereof, a COX inhibitor, a CDK4/6 inhibitor, a pan- CDK inhibitor, and combinations thereof.

The method of embodiment 11, wherein the senolytic p53/pRb activator comprises a nutlin, and wherein the nutlin is nutlin-3A. The method of embodiment 11, wherein the senolytie p53/pRb activator comprises a COX inhibitor, and wherein the COX inhibitor is selected from the group consisting of aspirin, ibuprofen, naproxen, diclofenac, indomethacin, and combinations thereof.

The method of embodiment 13, wherein the senolytie p53/pRh activator comprises diclofenac.

The method of embodiment 11, wdierein the senolytie p53/pRb activator comprises a pan CDK inhibitor, and wherein the pan CDK inhibitor is selected from the group consisting of flavopiridol, olomoucine II, purvalanol A, SNS-032, dinaciclib, MK-7965,

SCH727965, AT7519, R547, AZD5438, and AG024322.

The method of embodiment 11, wherein the senolytie p53/pRb activator comprises a CDK4/6 inhibitor, and wlierein the CDK4/6 inhibitor is selected from the group consisting of ribociclib, palbociclib, abemaciclib, and trilaciclib.

The method of any one of embodiments 1-16, wdierein the NAD+ supplement comprises Nicotinic Acid (NA), Nicotinamide Riboside (NR), Nicotinamide Mononucleotide (NMN), and/or Nicotinamide (NAM).

The method of embodiment 17, wherein the NAD+ supplement comprises: (a) NA and (b) one or more of NR, NMN and NAM.

The method of embodiment 17, wherein the NAD+ supplement comprises NR and NA. The method of any one of embodiments 1-19, wherein the insulin sensitizer, the mTQR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytie p53/pRb activator and/or the NAD+ supplement are nutraceutical agents. The method of any one of embodiments 1-20, wherein the aging-related disease or condition is selected from the group consisting of cancer, cardiovascular diseases, central nervous system diseases, Alzheimer's disease, Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, osteoporosis, arthritis, insulin-resistance and type II diabetes, benign prostatic hyperplasia, systemic lupus erythematosus, auto-immune diseases, psoriasis, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer, eczema, pain, inflammation or strain of muscle, tendon or joint, lipolysis, post !ipectomy and liposuction, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis, arthritis, and combinations thereof. The method of any one of embodiments 1-20, wherein the method is for cosmetic treatment.

The method of any one of embodiments 1 -20, wherein the aging-related disease or condition is obesity, non-alcoholic fatty liver disease, non-alcoholic steatotie hepatitis, pre-diabetic, diabetes type II and other fat or inflammatory' induced metabolic imbalances ( e.g , hypercholesterolemia).

The method of any one of embodiments 1 -23, wherein the individual further receives a fat reduction by lipectomy or modification by a lipoly sis procedure (such as the surgical procedure of liposuction and/or application of cold, heat, laser, ultrasound,

radiofrequency ablation for lipolysis and fat sculpturing).

The method of any one of embodiments 1-24, wherein the insulin sensitizer, the mTQR inhibitor, the agent having dual functions as an insulin sensitizer and an mTQR inhibitor, the senolytic p53/pKb activator, and/or the NAD+ supplement is administered systemically.

The method of any one of embodiments 1-24, wherein the insulin sensitizer, mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTQR inhibitor, the senolytic p53/pRb activator and/or the NAD+ supplement is administered via a combination of local and systemic routes.

The method of any one of embodiments 1-24, wherein the insulin sensitizer, the mTQR inhibitor, the agent having dual functions as an insulin sensitizer and an mTQR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered locally. The method of any one of embodiments 5-8 and 11-27, wherein the effective amount of rapamyein or the rapalog maintains a trough blood level of rapamycin selected from no more than about 8ng/ml for systemic administration, no more than about 2ng/ml for local administration, and no more than about IQng/ml for combined systemic and local administration, or an equivalent trough blood level of the rapalog thereof.

The method of embodiment 27 or 28, wherein the aging-related disease or condition is selected from the group consisting of primary or metastatic cancer lesions,

cardiovascular diseases, central nervous system diseases, Alzheimer's disease,

Parkinson's disease, age-related macular degeneration, cataract, retinopathy, skeletal muscular diseases, aging skin pigmentation, seborrheic keratosis, actinic keratosis, hair loss, alopecia, photo-aged skin, wrinkles, skin spots, skin cancer, psoriasis, eczema, pain. inflammation or strain of muscle, tendon or joint, lipofysis, post lipectomy and liposuction, non-alcoholic faty liver disease, non-alcoholic steatotic hepatitis, and combinations thereof.

The method of any one of embodiments 27-29, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement is administered via an implanted device

The method of embodiment 30, wherein the implanted device is selected from the group consisting of an intravesical or other urogenital tract gel, an intra-cranial or intra-thecal device, intra-ocular device, a drug-eluting stent, and CT, MRi or X-ray guided and inserted devices

The method of any one of embodiments 1-31, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAEM- supplement is formulated in a pharmaceutical composition comprising a pharmaceutically acceptable excipient.

The method of embodiment 31 or 32, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and the NAD+ supplement are formulated in a single pharmaceutical composition.

The method of embodiment 31 or 32, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and/or the NAD+ supplement are formulated in two or more pharmaceutical compositions.

The method of embodiment 34, wherein the insulin sensitizer, the mTOR inhibitor, the agent having dual functions as an insulin sensitizer and an mTOR inhibitor, the senolytic p53/pRb activator, and the NLO · supplement are each formulated in a different matrix for immediate or extended release.

The method of any one of embodiments 32-35, wherein the pharmaceutically acceptable excipient is a serum, gel, buffer solution, cream, lotion, liniment, ointment or

combinations thereof

The method of any one of embodiments 32-36, wherein the insulin sensitizer or the agent having dual functions as an insulin sensitizer and an mTOR inhibitor is in a pharmaceutical composition comprising at least about 0.5% (e.g., about 0.5-10%, w/v) metformin.

The method of any one of embodiments 32-37, wherein the insulin sensitizer is in a pharmaceutical composition comprising at least about 0.03% (w/v) piogiitazone.

The method of any one of embodiments 32-38, wherein the mTQR inhibitor is in a pharmaceutical composition comprising at least about 0.01% (w/v) rapamycin.

The method of any one of embodiments 32-39, wherein the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 1% (w/v) aspirin.

The method of any one of embodiments 32-39, wherein the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0.5% (e.g., about 0.5-4%, w/v) diclofenac.

The method of any one of embodiments 32-39, wherein the senolytic p53/pRb activator is in a pharmaceutical composition comprising at least about 0.3% (w/v) nutlin-3.

The method of any one of embodiments 32-42, wherein the NAD+ supplement is in a pharmaceutical composition comprising: (a) at least about 0.5% (w/v) Nicotinic Acid, and (b) at least about 0.5% (w/v) Nicotinamide Riboside (NR), at least about 1.5% (w/v) Nicotinamide Mononucleotide (NMN), and/or at least about 0.5% (w/v) Nicotinamide (NAM).

The method of any one of embodiments 32-42, wherein the NAD+ supplement is in a pharmaceutical composition comprising at least 0.5 % (e.g., about 0.5-6%, w/v) NR and/or at least 0.5 % (e.g., about 0.5-6%, w/v) NA

The method of any one of embodiments 27-44, wherein the aging-related disease or condition is an allergic or auto-immune skin disease, and wherein the method further comprises locally administering an effective amount of a corticosteroid to the individual. The method of any one of embodiments 1-45, wherein the method further comprises administering to the individual an effective amount of vitamin D, vitamin E, vitamin A, resveratrol, pterostilbene, L-carnitine, R-lipoic acid, leptine, a caspase inhibitor, and/or Chayote (Sechium edule) juice or extract.

The method of embodiment 46, wherein the method further comprises administering to the individual pterostilbene in a pharmaceutical composition comprising at least 0.5 % (e.g, about 0.5-3%, w/v) pterostilbene. The method of any one of embodiments 1-47, wherein the effective amount of the NAD+ supplement maintains a blood level of at least about 500mM NAD+.

The method of any one of embodiments 1-48, wherein the individual is a human.

The method of any one of embodiments 1-48, wherein the individual is a pet.

A method of decelerating, stabilizing, ameliorating, or reversing one or more signs of aging in a mammalian cell, comprising contacting the mammalian cell with:

(i) an effective amount of an insulin sensitizer,

(ii) an effective amount of an mTOR inhibitor,

(iii) an effective amount of a senolytic p53/pRb activator, and

(iv) an effective amount of an NAD+ supplement.

A method of decelerating, stabilizing, ameliorating, or reversing one or more signs of aging in a mammalian cell, comprising contacting the mammalian cell with:

(i) an effective amount of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) an effective amount of a senolytic p53/pRb activator, and

(iii) an effective amount of an NAD+ supplement.

A pharmaceutical composition comprising:

(i) an insulin sensitizer,

(ii) an mTOR inhibitor,

(iii) a senolytic p53/pRb activator, and

(iv) an NAD+ supplement.

A pharmaceutical composition comprising:

(i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) a senolytic p53/pRb activator, and

(iii) an NAD+ supplement;

and a pharmaceutically acceptable excipient.

A pharmaceutical composition comprising about 0.5-10% (w/v) metformin, about 0.5-4% (w/v) diclofenac, about 0.5-6% (w/v) nicotinamide riboside, about 0.5-6% (w/v) nicotinic acid, about 0.5-10% (w7v) pterostilbene, and a pharmaceutically acceptable excipient.

The pharmaceutical composition of embodiment 55, wherein the pharmaceutical composition is formulated for topical administration. 57. The pharmaceutical composition of embodiment 55 or 56, further comprising about 1-10% (w/v) urea, about 1-10% (w/v) sodium acetate, about 1-30% (w/v) ethanol, about 5-40% (w/v) PEG200, and about 5-40% (w/v) PEG400.

58. A kit for treating or preventing an aging-related disease or condition, comprising:

(i) an insulin sensitizer,

(ii) an mTOR inhibitor,

(iii) a senolytic p53/pRb activator, and

(iv) an NAD+ supplement.

59. A kit for treating or preventing an aging-related disease or condition, comprising:

(i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) a senolytic p53/pRb activator, and

(iii) an NAD+ supplement.

60. An implantable device for treating or preventing an aging-related disease or condition, comprising:

(i) an insulin sensitizer,

(ii) an mTOR inhibitor,

(iii) a senolytic p53/pRb activator, and

(iv) an NAD+ supplement.

61. An implantable device for treating or preventing an aging-related disease or condition, comprising:

(i) an agent having dual functions as an insulin sensitizer and an mTOR inhibitor;

(ii) a senolytic p53/pRb activator, and

(iii) an NAD+ supplement.

EXAMPLES

[0173] The examples below are intended to be purely exemplary of the invention and should therefore not be considered to limit the invention in any way. The following examples are offered by way of illustration and not by way of limitation.

Clinical studies of combination regimens

[0174] The invention is based on preliminary data from clinical trials of two exemplary anti aging combination regimen as described below. [0175] The clinical trials (A-C) were interventional, Phase I/II open label, studies of two combination regimens for promoting healthy aging in human individuals.

[0176] The first clinical trial (“systemic Trial A”) studied a four-component combination regimen consisting of an insulin sensitizer, a senolytic p53 activator, an mTOR inhibitor, and a NAD · supplement, which was administered systemically. The interventions received by the individuals included: (1) insulin sensitizer: metformin at 500mg twice a day after meals on the seventh week followed by an escalation to 750mg twice a day after meals on the eighth week onwards, (2) senolytic p53 activator: ibuprofen at a dose of 200m g twice a day for 3 days/2 weeks in the first week followed by an escalation to 4Q0mg twice a day for 3 days/2 weeks from the second week onwards; (3) mTOR inhibitor: rapamycin at 3 mg/2 weeks, given on the first day of the ibuprofen dose every 2 weeks; and (4) NAD+ supplement: (a) niacin at 125 mg twice a day on the third week followed by an escalation to 250 rng twice a day on the fourth week omvards; and (b) nicotinamide riboside (NR) at 750 mg twice a day or the fifth w^eek followed by an escalation to lOOOmg twice daily on the sixth week onwards. Rapamycin was stopped on the 14 th w ? eek and an office examination and laboratory assays in the same week. All other medications such as ibuprofen (bid for 2 days/week), nicotinic acid (NA) and nicotinamide riboside (NR), and metformin twice daily continued on the 14 th week. All medications were resumed as of the 15 week for 6 more weeks (second cycle). Then, rapamycin alone was stopped in the 7 th week in this second cycle. Subsequent treatment cycles are the same: 6 w'eeks rapamycin on and 1 week off, accompanied by examination and laboratory assay in the 7 th week.

[0177] The second clinical trial (“local Trial B”) studied a three-component combination regimen consisting of an agent having dual functions as an insulin sensitizer and an mTOR inhibitor, a senolytic p53 activator, and a NAD+ supplement in treating osteoarthritis, which was administered topically. The interventions received by the individuals included: a serum

(“LASO”) consisting of an aqua/PEG 200/PEG400/ethanol emulsion of diclofenac 2%, metformin 2%, nicotinic acid 1.5%, nicotinamide riboside 3%, and pterostilbene 1.5%. A phase I three-step escalating modified Fibonacci study involved groups of three patients being treated with either diclofenac 2% serum, followed by a washout period of two weeks before a metformin 2% serum, and then after a second washout period of two weeks, to start finally the LASO serum. The serum was applied to locally to a kno n inflamed, or stiffness, or painful joint site in the peripheral joints such as the knees, fingers, wrist or the spine. [0178] The third clinical trial (“local Trial C”) studied the efficacy of the LA80 serum in promoting post iipolysis fat homeostasis, and treating diabetic type 2 glucose resistance, non- alcoholic fatty liver disease, non-alcoholic steatotic hepatitis for post Iipolysis fat homeostasis, diabetic type 2 glucose resistance, non-alcoholic fatty liver disease, non-alcoholic steatotic hepatitis LASO was applied in groups of three patients in three escalating steps as a modified Fibonacci study. The target was abdominal fat. LASO was applied up to 15 cc onto the abdominal fat twice daily followed by a massage.

[0179] Healthy individuals of either gender and at least 50 years old and no history of diabetes (Type 1 or Type 2), and having an Hbalc (Hemoglobin A1C) level of 6.4 or less were eligible to enroll in the study. About 20 individuals are targeted to be enrolled in the study. These inclusion criteria were chosen to study subjects who may have evidence of impaired glucose regulation, but are not yet diabetic.

[0180] Exclusion criteria for this clinical trial included: (!) serious chronic or acute illness, including cancer, clinically significant congestive heart failure, COPD, inflammatory conditions, serum creatinine > 1.4 mg/dl (female) or > 1.5 mg/di (male), active liver disease, history of metabolic acidosis, poorly controlled hypertension, epilepsy, recent (within 3 months) CVD event (Ml, PTC A, CABG, stroke), history of bariatric or other gastric surgery, cigarette smoking, binge alcohol use (>7 drinks in 24 hrs); (2) treatment with drugs known to influence insulin sensitivity and/or immune functions (other diabetes medications, systemic glucocorticoids); and hypersensitivity to any component of the treatment or formulation.

[0181] The primary outcome measures the safety of the combination regimen, as assessed at 14 weeks, 28 weeks and 1 year from the inception of the treatment. This outcome addresses safety concerns related to long-term inhibition of mTOR by administration of rapamycin or metformin, with or without a nutraceutical agent. The primary outcome measures of local Trial B also include the efficacy evaluated by WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) at 14 and 28 weeks. The primary outcome measures of local Trial C include efficacy of fat weight reduction, both subcutaneous and visceral wise as assessed by a DEXA scan, as well as insulin sensitivity index such as fasting glucose, Hbalc or HQMA-IR, liver function tests and IL6 as an inflammatory markers.

[0182] The secondary outcomes include the following; (1) assessment of insulin sensitivity and insulin secretion at 14, 28 weeks and every 6-7 months afterwards post treatment by C peptide, fasting glucose and Hbalc; (2) determination of telomere length at week 14 and at 6-7 months afterwards post treatment and then yearly afterwards; (3) determination of IL6 level at week 14 and t 6-7 months afterwards post treatment and then yearly afterwards; and (4) assessment of rates of aging-related diseases at week 14 and at 6-7 months afterwards post treatment and then yearly afterwards. Aging-related diseases assessed in this experiment include cancer, cardiovascular, central nervous system diseases (e.g., Alzheimer’s), and metabolic diseases (e.g., type 2 diabetes). The rates of incidence of aging-related diseases in individuals treated with the combination regimen in the long-term (e.g., at the 10 year mark) may show a decrease compared with historic controls; (5) Insulin sensitivity is estimated from insulin and glucose levels obtained following the standard meal challenge, using HOMA-IR, or a modification of the Matsuda index, or with triglycerides level and glucose/insulin level. (6) Telomere length in cells from PBMC or oral mucous membrane samples from individuals treated with the combination regimen is determined. The telomere length may be stabilized, shortened, or prolonged in response to the combination regimen and aging. The assay used is Tel o Year and telomere lengths are measured at week 14, and at 6-7 months afterwards post treatment and then yearly afterwards; (7) The IL6 level is a good measurement of inflammation related to aging, which may stabilize, decrease or increase in response to the combination regimen. The 1L6 level is measured at week 14, and at 6-7 months afterwards post treatment and then yearly afterwards; (8) Fat stored as subcutaneous, visceral adipose tissue and muscle mass at w¾ek 14, and at 6-7 months afterwards post treatment and then yearly afterwards; (9) Liver functions assay, albumin level and eGFR are also measured accordingly.

[0183] Investigational outcome measures assess the efficacy of the combination regimen in individuals having different levels of glucose regulation capacity. Individuals are classified into three groups: those with Hbalc level of 5 6 or below, those with Hbalc level of 5.7 to 6.0, and those with Hbalc of 6.1 to 6.4. The insulin sensitivity outcome as defined by HOMA-IR, 11.6 level, telomere length and NAD+ (metabolomes) levels of the three groups of individuals are compared. Individuals with mTOR activation may have more insulin resistant cells driven into irreversible senescence so that mTOR inhibition has less anti-aging effect when combined with senolytic p53 therapy. This effect may be more apparent in individuals having a high insulin resistant factor to start with, such as those with a HOMA-IR over 2.

[0184] Preliminary results at the time of filing have demonstrated safety of the combination regimen in both systemic as well as in the local trials. As of this filing, a total of 15 patients were enrolled in the studies. The only toxicides reported (10%) for the systemic Trial A were all limited to grade 1-2, such as abdominal discomfort, decreased appetite, diarrhea and some minor muscle pain or cramping. The only toxicities reported (23%) for the local Trial B and C were also limited to grade 1-2 skin sensitivity symptoms such as redness, itchiness and minor muscle ache.

[0185] Table 1 and FIG. 1 show results from Patient 1 on the systemic Trial A, who has been treated with the combination regimen for over 1 year.

Table 1. Patient 1 data from 2017-2018 in systemic Trial A.

[0186] Table 2 and FIG. 2 show results from Patient 2 on the systemic Trial A, who has been treated with the combination regimen for over 1 year.

Table 2. Patient 1 data from 2017-2018 in systemic Trial A.

[0187] Table 3 below shows results from 11 patients on local Trial B.

Table 3. Patient Characteristics in local Trial B.

0188] Table 4 and FIG. 3 show results from Patient 1 on the local Trial C, who has been treated with the combination regimen from April 2018 to October 2018.