Delivery Strategies of siRNA Therapeutics for Hair Loss Therapy

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This is a comprehensive review describing the development of siRNA based therapeutics for hair loss. Overall the manuscript is organized well and easy to follow. However, I have some below suggestions for improvement:

1, the title is not easy to understand. Please revise and make it easy to comprehend. 

2, please include the dose investigated for siRNA therapeutics.

3, please provide a paragraph or table comparing the targets of siRNA for hair loss. 

4, It seems the clinical investigation of siNRA for hair loss is still limited. Please explain why. 

Author Response

Re: Response to Comments and Suggestions for Authors

 

Manuscript ID: ijms-3015952

Type of manuscript: Review

Title: Delivery strategies of siRNA therapeutics for hair loss overcoming barriers based on clinical impact

 

June 24, 2024

 

Dear Editor-in-chief,

 

                We are pleased to submit our responses to the reviewers’ comments and suggestions. We answered the reviewers’ questions one by one after considerable discussion. The revised manuscript has been corrected and modified according to the reviewers’ comments and the changes in the revised manuscript are presented in red. We hope the revised manuscript to meet your high standards for publication. Thank you for your consideration.

 

Reviewer #1

This is a comprehensive review describing the development of siRNA-based therapeutics for hair loss. Overall the manuscript is organized well and easy to follow. However, I have some below suggestions for improvement:

 

1. The title is not easy to understand. Please revise and make it easy to comprehend.

Thank you for your comment. We modified the title in the revised manuscript as follows: “Delivery strategies of siRNA therapeutics for hair loss therapy”. This manuscript describes siRNA delivery systems for hair loss therapy that overcome intrinsic and extrinsic barriers. We analyzed siRNA delivery systems in previously approved siRNA products and delivery systems for hair loss therapy in clinical trials, which can be repurposed for siRNA therapeutics in hair loss therapy. The siRNA delivery systems developed in current approaches are also discussed based on delivery strategies for siRNA therapeutics in hair loss therapy.

Page on 1: Title

Delivery strategies of siRNA therapeutics for hair loss therapy

Su-Eon Jin* and Jong-Hyuk Sung*

Epi Biotech Co., Ltd., Incheon, Republic of Korea

 

2. Please include the dose investigated for siRNA therapeutics.

                Thank you for your comment. The investigated dose of siRNA therapeutics is a key factor in clinical use. In Table 1 of the revised manuscript, siRNA dose levels were additionally added in non-clinical/clinical models as follows. They ranged at 6 μg to 1 mg for topical and intradermal applications. Delivery systems are introduced to prolong the effects of siRNA.

 

Pages on 30-31: Table 1

Table 1 Current non-clinical/clinical approaches to siRNA therapeutics in hair loss.

Target siRNA (sequences)	Nanoplatform for delivery	siRNA dose	Outcome	Reference
CXCL12 siRNA (5’-GAACAACAACAGACAAGUG-3’, 3’-CUUGUUGUUGUCUGUUCAC-5’)	-	6 μg (subcutaneous injection, every 2 days), C3H/HeN mice; 1 μg, hair organ culture	Triggering telogen-to-anagen transition Increasing hair length in hair organ culture	[64]
T-box21 siRNA (5’-UGAUCGUCCUGCAGUCUCUdTdT-3’, 3’-dTdTACUAGCAGGACGUCAGAGA-5’)	Cationized gelatin conjugation Cationized gelatin microsphere (controlled delivery)	10 μg (subcutaneous injection), C3H/HeJ mice, alopecia areata model	Restoring hair shaft elongation	[65]
CXXC5 siRNA	-	10 μM, HaCaT	Recovering hair growth suppressed by PGD2	[66]
EGLN1 or EGLN3 siRNA SFRP2 or SERPINF1 siRNA	Lipofectamine RNAi MAX transfection reagent	0.04 nmol/L (2-day transfection), human hair follicle culture	Promoting dermal papilla proliferation and hair follicle growth with prolonged anagen stage and delayed catagen transition	[67]
FGF5 or FGF18 siRNA	Cholesterol conjugation Cream	20 μM (50 μL, intradermal injection or topical application), C57BL/6 mice, healthy model	Restoring hair growth	[68]
Androgen receptor siRNA (SAMiRNA)	Polyethylene glycol (PEG) and hydrophobic hydrocarbon conjugates at each end of unmodified DNA/RNA heteroduplex (99.2 ± 5.1 nm (22°C, 55% ± 5 humidity) and 105.0 ± 2.5 nm (40°C, 75% ± 5 humidity)) Hair tonic (ethanol (15%, v/v), niacinamide (1% w/v), betaine (1% w/v), biotin (0.02% w/v) and buffer in aqueous solution) (0.5 mg/mL and 5 mg/mL)	Androgenetic alopecia, clinical study 0.5 mg/mL three times per week: 45, male (test article 8; placebo 6) and female (test article 14; placebo 17); 5 mg/mL once a week: 43, male (test article 9; placebo 10) and female (test article 13; placebo 11))	Increasing total hair counts after administrating for 24 weeks	[80]
Androgen receptor asymmetric siRNA (asiRNA)	Cholesterol conjugation, chemical modification	1.0 μM, human dermal papilla cells; 3 μM and 6 μM, ex vivo human hair follicle culture; 0.125 mg, 0.25 mg, 0.5 mg, and 1.0 mg (intradermal injection), C57BL/6 mice; 0.125 mg, 0.25 mg, and 0.5 mg (intradermal injection), androgenetic alopecia mouse model (dihydrotestosterone daily injection, 25 mg/kg)	Decreasing telogen propagation and increasing the mean hair bulb diameter; Attenuating dihydrotestosterone-mediated increases in interleukin-6, transforming growth factor-β1, and dickkopf-1 levels; No significant toxicity	[70]

 

 

 

3. Please provide a paragraph or table comparing the targets of siRNA for hair loss. 

                Although the siRNA targets for hair loss therapy were presented in section “4.2. siRNA targets for hair loss” and Table 1 in the revised manuscript, the targeting approaches were described in greater detail.

 

Pages on 12: 4.2. siRNA targets for hair loss

~Hair loss-associated signaling pathways have been targeted to discover therapeutic siRNA, as scientifically evidenced by multiple markers of hair follicles used to identify the structure and cell composition due to their complexity [63]. Signaling pathways targeting hair loss are consistently connected to the regulation of hair growth cycle control of hair morphogenesis and regeneration. Among the pathways that cause hair loss, siRNA therapeutics have been studied for targeting C-X-C motif chemokine ligand 12 (CXCL12) [64], Th1 transcription factor  (T-box21) [65], CXXC finger protein 5 (CXXC5) [66], egg-laying-defective 9 (EglN) hypoxia inducible factor (HIF)-1 prolyl-hydroxylase (EGLN1 or EGLN3), secreted frizzled related protein 2 (SFRP2) or serpin family F member 1 (SERPINF1) [67], FGF5 or FGF18 [68], and AR [69, 70], with doses on the microgram-to-gram scale based on siRNA efficacy. These are summarized in Table 1, which displays delivery systems and experimental outcomes in non-clinical/clinical models for hair loss.~

Overall, the therapeutic development for hair loss is diversely and successfully challenged using siRNA in a multiangle approach based on the proof-of-concept of diseases, beyond AR targeting due to the off-target effect of DHT blockade [32]. The delivery paradigms of drug modification (e.g., functional group and targeting ligands), environment modification (e.g., formulation), and delivery system introduction (e.g., nanoparticles) for optimization should be evolved and repurposed for clinical translation (see “5.2. siRNA delivery principles for hair loss”) [17]. ~

 

Pages on 30-31: Table 1, the same as No.2 answer

 

4. It seems the clinical investigation of siRNA for hair loss is still limited. Please explain why. 

                Thank you for your comment. The mechanisms of action for the pathophysiology of hair loss have been investigated, but they are sophisticated and complicated. Although finasteride and minoxidil are available for androgenetic alopecia, and baricitinib was recently approved for alopecia areata, multiple signaling pathways are associated with hair loss therapy in clinics. Clinically tunable targets for siRNA are still a challenge in terms of intrinsic and extrinsic barriers to siRNA therapeutics. Repurposing is an unprecedented strategy for clinical investigation of siRNA therapeutics for hair loss in the development of target identification and delivery system design. We further discuss repurposing from this point of view. The revised manuscript contains this explanation as follows.

 

Pages on 6-7: 3. Clinical trials for hair loss

Clinical trials for hair loss have been searched in the clinicaltrials.gov database using hair loss as a keyword (assessed on March 26, 2023). The intervention of siRNA was undetected in the search results of clinical trials for hair loss so far because clinically tunable targets for siRNA are still a challenge in terms of intrinsic and extrinsic barriers to siRNA therapeutics. However, topical formulations or nanoplatforms displaying potential clinical impacts have been introduced to improve the hair loss conditions even in a combination therapy. Repurposing is an unprecedented strategy for clinical investigation of siRNA therapeutics for hair loss in the development of target identification and delivery system design. In particular, delivery systems from the searched clinical trials in hair loss can be repurposed for siRNA therapeutics after interchangeable modification followed by design principles depending on siRNA characteristics. Design principles and delivery systems overcoming critical barriers will be further discussed in the following sections (see “4. Small interfering ribonucleic acid (siRNA) as a therapeutic” and “5. siRNA delivery”).

 

Sincerely,

 

Su-Eon Jin, Ph.D.

Advisory member, Epi Biotech Co., Ltd.

E-mail: [email protected]

 

Jong-Hyuk Sung, Ph.D.

CEO, Epi Biotech Co., Ltd.

E-mail: [email protected]

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

In the manuscript submitted for review, the authors have reviewed the literature on the characterisation of siRNA delivery systems, with a view to their potential use in hair loss therapy. The topic addressed by the authors is important and timely. The manuscript is clearly written and presented in a well-organised manner. The literature cited is mostly recent and relevant publications. Figures and tables are appropriate. Conclusions are consistent with the evidence and arguments presented. Although the article contains enough novelty, I suggest several improvements:

1. In my opinion, the article would have benefited if the authors had formulated the research question and the inclusion and exclusion criteria for publications in the review.

2. Table 1 is too long. I suggest dividing it.

3. Could the authors discuss the current legal status governing the use of nanoparticles including siRNA nanoparticles in therapy?

4. "References" please prepare according to the authors' instructions.

Author Response

Re: Response to Comments and Suggestions for Authors

 

Manuscript ID: ijms-3015952

Type of manuscript: Review

Title: Delivery strategies of siRNA therapeutics for hair loss overcoming barriers based on clinical impact

 

June 24, 2024

 

Dear Editor-in-chief,

 

                We are pleased to submit our responses to the reviewers’ comments and suggestions. We answered the reviewers’ questions one by one after considerable discussion. The revised manuscript has been corrected and modified according to the reviewers’ comments and the changes in the revised manuscript are presented in red. We hope the revised manuscript to meet your high standards for publication. Thank you for your consideration.

 

Reviewer #2

 

In the manuscript submitted for review, the authors have reviewed the literature on the characterisation of siRNA delivery systems, with a view to their potential use in hair loss therapy. The topic addressed by the authors is important and timely. The manuscript is clearly written and presented in a well-organised manner. The literature cited is mostly recent and relevant publications. Figures and tables are appropriate. Conclusions are consistent with the evidence and arguments presented. Although the article contains enough novelty, I suggest several improvements:

 

1. In my opinion, the article would have benefited if the authors had formulated the research question and the inclusion and exclusion criteria for publications in the review.

                This manuscript describes siRNA therapeutics for hair loss therapy with repurposed siRNA designs, delivery systems in approved siRNA therapeutics, and clinically tunable formulations from clinical trials for hair loss. We firstly discussed the need for siRNA therapeutics in hair loss therapy and identified barriers to siRNA therapeutics, including siRNA design, currently defined siRNA targets, and delivery systems. Specifically, currently available approaches to target identification and formulation for siRNA were included. However, monogenic conjugation for siRNA delivery, which includes modifying siRNA structure, was excluded from the revised manuscript.

 

Page on 4: 1. Introduction

~ We introduce clinically impacted siRNA delivery systems for hair loss based on the design principles overcoming intrinsic and pathophysiological barriers beyond siRNA engineering. In this review, current clinical trials for hair loss are described regarding intervention, condition, phase and study design, grounded in an overview of hair loss. Designed delivery systems repurposed for therapeutic siRNA are also summarized from fundamental siRNA characteristics to optimized delivery after analyzing current non-clinical and clinical approaches in siRNA platform development for hair loss. We focus on formulation of nanocarriers rather than structural modification of siRNA. A strategic research framework for siRNA delivery systems in hair loss is further provided as a path toward enhanced performance.

 

2. Table 1 is too long. I suggest dividing it.

                Table 1 presents the delivery approaches from clinical trial data for hair loss. We analyzed the clinical trials registered to clinicaltrials.gov, categorized by intervention, dose/delivery system, administration route, and condition. Subcategories of interventions were included as (1) small molecules, corticosteroids and others, (2) Jak inhibitors and antibodies, (3) cell therapy or cell therapy-related products, (4) medical devices, and (5) formulations or delivery systems. Although this table provides useful information, it is too long to put in the revised manuscript. We made it a supplementary table in the revised manuscript according to the reviewer’s suggestion as follows.

 

Page on 8: 3.2. Primary interventions and delivery platforms for hair loss

The key interventions of clinical trials are enumerated to confirm hair loss therapy for prevention and treatment, in combination with feasible formulations or nanocarriers (Supplementary Table S1). They are classified by small molecules, corticosteroids and others [e.g., finasteride, dutasteride, minoxidil, methylprednisolone, triamcinolone acetonide, hydroxychloroquine, and ALRV5XR (shampoo)] [31, 32], Jak inhibitors and antibodies (e.g., ifidancitinib, baricitinib, deuruxolitinib, jaktinib, ritlecitinib, ruxolitinib [33], and tofacitinib), cell therapy or cell therapy-related products [e.g., human autologous hair follicle cells, autologous cultured dermal and epidermal cells, adipose-derived stem cell suspension [34], hair stimulating complex, GID SVF-2, conditioned media from umbilical cord blood-derived stem cell culture, lipoaspiration, autologous fat graft enriched with adipose-derived regenerative cells (ADRCs) [35], platelet-rich plasma (PRP) [36]], and medical devices [e.g., light therapy (Theradome LH80 pro, REVIAN 101), Derma pen (microneedle pen), Dermojet (needle-less syringe), DMEP kit (cryotherapy), MTS-01 (microneedle), scalp cooling (Paxman cooling machine), UV, Venus glow™ (skin renewal machine with serum), and HairDx (genetic testing for baldness)] in terms of pharmacology. Apart from targeting androgens in hair loss to simply remove, zinc supplement was also applied with a minoxidil solution (5%) [37].

 

Attachment: Supplementary Table S1

Supplementary Table S1. Delivery approaches based on key interventions in clinical trials for hair loss.

Category	Key intervention	Dose/ Delivery system	Administration route	Condition	NCT number
Small molecules, corticosteroids and others	Finasteride	1 mg	-	Androgenetic alopecia (female)	NCT01052870
	Dutasteride	0.02, 0.1 and 0.5 mg	Oral	Androgenetic alopecia	NCT01231607
		0.5 mg	Oral	Male pattern hair loss or androgenetic alopecia	NCT02014584
	Minoxidil	5%, q.d. (foam) 2%, b.i.d. (solution)	Topical	Female pattern hair loss	NCT01145625
		5% (90% ethanol and 5% propylene glycol)	Topical	Female pattern hair loss	NCT04090801
		5% (foam)	Topical	Female pattern hair loss	NCT01226459
		5% (noisome, spray)	Topical	Alopecia areata	NCT05587257
	Methylprednisolone (sodium succinate)	15 mg/kg (200 mL fresh orange juice)	Oral	Alopecia totalis Alopecia universalis Ophiasic alopecia	NCT01167946
	Triamcinolone acetonide	5 mg/mL	Intralesional	Alopecia areata	NCT03535233
	Hydroxychloroquine	-	-	Alopecia areata	NCT00176982
	ALRV5XR	1 each (capsule)	Oral	Androgenetic alopecia	NCT04450602
		3-7 mL (shampoo)	Topical	Telogen effluvium Hair thinning Hair loss/baldness
		1 each (capsule)	Oral	Androgenetic alopecia	NCT04450589
		3-7 mL (shampoo)	Topical	Telogen effluvium
		1 mL (serum)	Topical	Hair thinning Hair loss/baldness
	Zinc supplement with minoxidil solution (5%)	15 mg chelate zinc supplement: additional to 5% minoxidil (solution)	Oral	Female pattern hair loss	NCT01662089
Jak inhibitors and antibodies	Ifidancitinib (ATI-50002)	0.12 and 0.46% (solution)	Topical	Alopecia areata	NCT03354637
	Baricitinib (LY3009104)	2 and 4 mg (tablet)	Oral	Alopecia areata	NCT03899259
		2 and 4 mg (tablet)	Oral	Severe or very severe alopecia areata	NCT03570749
		High and low (tablet)	Oral	Alopecia areata Alopecia Hypotrichosis Hair diseases Skin diseases	NCT05723198
	Deuruxolitinib (CTP-543)	8 and 12 mg (tablet)	Oral	Alopecia areata	NCT04797650
	Jaktinib	50 and 75 mg (tablet)	Oral	Alopecia areata	NCT05051761
		50, 150 and 200 mg (tablet)	Oral	Alopecia areata	NCT04034134
		0.5, 1.5 and 2.5% (cream)	Topical	Alopecia areata	NCT04445363
	Ritlecitinib (PF-06651600)	200 mg (q.d., 8 weeks) and then 100 mg (q.d., 40 weeks) (tablet)	Oral	Cicatricial alopecia	NCT05549934
	Ruxolitinib	20 mg (tablet)	Oral	Alopecia areata	NCT01950780
		0.6% (cream)	Topical	Alopecia areata	NCT02553330
		Initial dose and maintenance dose	Oral	Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED) Alopecia areata	NCT05398809
	Tofacitinib	5 – 10 mg	Oral	Alopecia areata	NCT02299297
Cell therapy or cell therapy-related products	Human autologous hair follicle cells	-	Injection to scalp	Androgenetic alopecia	NCT01286649
	Autologous cultured dermal and epidermal cells with 5% minoxidil	-	Injection to scalp	Androgenetic alopecia Male pattern baldness Female pattern baldness	NCT01451125
	Adipose derived stem cell suspension plus platelet rich plasma	-	Follicular unit extraction	Androgenetic alopecia	NCT03388840
	Hair stimulating complex	0.1 mL (30-gauge needle)	Intradermal injection to scalp	Androgenetic alopecia	NCT01501617
	Adipose-derived stromal vascular fraction (GID SVF-2)	-	Intradermal injection to scalp	Androgenetic alopecia	NCT02626780
	Conditioned media of umbilical cord blood-derived stem cells (NGF-574H)	Hair serum with 5% conditioned media of umbilical cord blood-derived stem cells	Directly used by subjects themselves at home	Androgenetic alopecia	NCT03676400
	Biocellular-cellular regenerative mixture	-	Intravenous infusion	Alopecia areata Scarring alopecia	NCT03078686
	Autologous fat graft enriched with adipose-derived regenerative cells (ADRCs)	Purified adipose + 500,000 or + 1,000,000 ADRCs	Subcutaneous injection in scalp	Androgenetic alopecia	NCT02503852
	Platelet-rich plasma	6 mL, activated with or without pulsed electrical fields	Subcutaneous injection	Androgenetic alopecia	NCT05348343
		-	Injection to half head	Androgenetic alopecia	NCT02074943
		-	Injection to scalp	Androgenetic alopecia	NCT03376581
Medical devices	Theradome LH80 pro for photobiomodulation therapy combined with scalp cooling	A wearable laser helmet device, thrice weekly	Light therapy to scalp	Chemotherapy-induced alopecia	NCT05177289
	REVIAN 101	A cap for portable use with rechargeable battery and adapter with active LEDs, a daily 10-minute treatment over the course of 26-weeks	Modulated light therapy to scalp	Androgenetic alopecia	NCT04019795
	Derma pen	Microneedling combined with methotrexate (25 mg/mL) at 0.02 mL/cm2, a maximum of 0.1-0.2 mL (2.5-5 mg)	Topical	Alopecia areata	NCT05485571
		Microneedling after 5% minoxidil application	Topical	Alopecia areata	NCT05587257
	Dermojet	A needless syringe, 0.1 mL of Depo-Medrol (methylprednisolone acetate) 40 mg/2 mL	Injection to scalp	Alopecia areata	NCT01017510
	Dimethyl ether and propane (DMEP) kit	Superficial cryotherapy using DMEP at -57℃	Topical	Alopecia areata	NCT04680234
	MTS-01	7% (gel in 100-mL tube)	Topical	Radiotherapy-induced alopecia	NCT00713154
	Paxman cooling machine	-	Topical	Chemotherapy-induced alopecia	NCT01008774
	UV	Narrow band UVB (311 nm) phototherapy	Topical	Alopecia areata	NCT03847441
		UVB excimer light, twice weekly	Topical	Alopecia areata	NCT01802177
	Venus glow™	Venus Glow hydradermabrasion device (cleansing and micromassaging)	Hydradermabrasion on the scalp	Androgenetic alopecia	NCT05426629
	HairDx	Sample collection kit for DNA	Saliva collection	Hair loss Hair loss/baldness Female pattern baldness Androgenetic alopecia	NCT04379583
Formulations or delivery systems	Aldara (imiquimod)	5% (cream)	Topical	Alopecia areata	NCT00177021
	Crisaborole	Ointment	Topical	Alopecia areata	NCT04299503
	Diphenylcyclopropenone (DPCP)	Ointment	Topical	Alopecia areata	NCT03651752
	LEO 124249	Ointment	Topical (eyebrow)	Alopecia areata	NCT03325296
	CU-40101	Liniment	Topical	Androgenetic alopecia	NCT05380427
	CU-40102	0.25% (2.275mg/mL) (spray)	Topical	Androgenetic alopecia	NCT05135468
	XN-001	Nitric oxide gel [14.6 mM sodium nitrite in distilled water with HEC1 (m.w. 50,000-1,250,000); 14.6 mM maleic acid and 14.6 mM ascorbic acid in distilled water with HEC]	Topical	Androgenetic alopecia	NCT01347957
	Targretin (bexarotene)	1% (gel)	Topical	Alopecia areata	NCT00063076
	Latanoprost	0.005% (solution)	Topical	Alopecia areata	NCT02350023
		Ophthalmic solution	Topical (eyelash)	Alopecia areata	NCT00187577
	Autologous platelet-rich fibrin matrix (PRFM)	0.1 mL (4-8 mL, isolated from 9-18 mL of peripheral blood)	Intradermal injection to scalp	Alopecia	NCT01590238
	Sodium valproate	Nanospanlastic dispersion	Topical	Alopecia areata	NCT05017454
	Nanofat	Autologous nanofat grafting	Injection to scalp	Androgenetic alopecia	NCT03506503
	Exosomes	100e10 particle (exosomes isolated from human amniotic mesenchymal stem cells)	Injection	Alopecia Hair loss (prevention)	NCT05658094

¹ Hydroxyethylcellulose

 

 

3. Could the authors discuss the current legal status governing the use of nanoparticles including siRNA nanoparticles in therapy?

                Thank you for your comment. The reference entitled “The regulation of nanomaterials and nanomedicines for clinical application: current and future perspectives” (Rachel Foulkes et al. Biomater. Sci., 2020, 8, 4653-4664. DOI: 10.1039/D0BM00558D.) describes the regulatory needs for nanoparticle products. The safety and efficacy of nanoparticle or nanoparticle-based products are at issue based on their clinical needs, administration route, and physiology because “one size does not fit all”. Nanoparticle usage for siRNA therapeutics in clinics should be guided by “Drug Products, Including Biological Products, that Contain Nanomaterials (FDA, 2022)” or “Considering Whether an FDA-Regulated Product Involves the Application of Nanotechnology (FDA, 2014)” published by the FDA. We additionally added the current legal status for nanoparticles in the revised manuscript as follows.

 

Page on 20: 5.1.2. Risk mitigation of siRNA therapeutics for hair loss

~ Overall, the safety and efficacy of nanoparticle or nanoparticle-based products are at issue based on their clinical needs, administration route, and physiology because “one size does not fit all” [97]. Although their complexed size, structure, and properties are still underestimated in the physiological environment followed by no ultimate clarification, nanoparticle characterization and usage for siRNA therapeutics in clinics should be guided by “Drug Products, Including Biological Products, that Contain Nanomaterials” [98] or “Considering Whether an FDA-Regulated Product Involves the Application of Nanotechnology” [99], published by the FDA. Characterization of the adequacy and complexity of structure and function, the mechanism of action for biological effects, in vivo release, in vitro-in vivo correlation, stability, nanotechnology maturity, and manufacturing changes is recommended to assess nanomaterials based on their engineered dimension, structure and function. ~

 

References

97. Moon, I. J.; Yoon, H. K.; Kim, D.; Choi, M. E.; Han, S. H.; Park, J. H.; Hong, S. W.; Cho, H.; Lee, D. K.; Won, C. H., Efficacy of Asymmetric siRNA Targeting Androgen Receptors for the Treatment of Androgenetic Alopecia. Mol Pharm 2023, 20(1), 128-135.
98. FDA, Drug Products, Including Biological Products, that Contain Nanomaterials. 2022.
99. FDA, Considering Whether an FDA-Regulated Product Involves the Application of Nanotechnology. 2014.

 

4. "References" please prepare according to the authors' instructions.

                We double-checked the cited references according to the authors’ instruction. In the revised manuscript, the references are as follows.

 

Pages on 30-51: References

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Sincerely,

 

Su-Eon Jin, Ph.D.

Advisory member, Epi Biotech Co., Ltd.

E-mail: [email protected]

 

Jong-Hyuk Sung, Ph.D.

CEO, Epi Biotech Co., Ltd.

E-mail: [email protected]

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors have addressed my concerns.