CYCLIC PEPTIDE AND A MEDICAMENT, EXTERNAL PREPARATION AND COSMETIC COMPRISING SAID CYCLIC PEPTIDE

Abstract

The present invention is aimed for providing a novel peptide with a high drug efficacy and strong effect, a medicament or external preparation comprising it, specifically a prophylactic or therapeutic for dermatitis, rhinitis or alopecia, or a hair growth stimulant, a hair growing agent, an antipruritic or a skin-care product. The present invention achieved said aim by providing a cyclic peptide having an amino acid sequence expressed by the Formula I or a derivative thereof or a pharmaceutically acceptable salt thereof, wherein the amino acid sequence does not have a peptide bond that is not between the amino acids constituting the amino acid sequence.

Claims

1.-15. (canceled)

16. A method of using an external preparation comprising applying the external preparation to skin and/or mucosa of a subject, wherein the external preparation comprises a cyclic peptide having an amino acid sequence expressed by the Formula I: ##STR00005## wherein, X.sup.1 denotes Gly, Val, Ala, Ser or Thr, X.sup.2 denotes Arg, Gln or His, X.sup.3 denotes Lys or Arg, X.sup.4 denotes Met, Leu or Ile, X.sup.5 denotes Be or Val, X.sup.6 denotes Ser or Gly, X.sup.7 denotes Ser or Ala, X.sup.8 denotes Ser, Gln, Val, Ala, Thr, Leu, Ile or Met, X.sup.9 denotes Ser, Val, Ala or Thr, X.sup.10 denotes Gly or Arg, X.sup.11 denotes Leu, Met, Ile, Val or Ala, X.sup.12 denotes Gly, Ser or Ala, and the line connecting two Cys denotes a disulfide bond, and wherein said amino acid sequence does not have a peptide bond that is not between the amino acids constituting said amino acid sequence, or a derivative thereof or a pharmaceutically acceptable salt thereof.

17. The method of using the external preparation according to claim 16, wherein the mucosa is labial, oral, nasal, ocular or vaginal mucosa.

18. The method of using the external preparation according to claim 16 for treating and/or preventing dermatitis, for alleviating or resolving itch, for treating eczematous or other erosion or ulcer, or for skin-care.

19. The method of using the external preparation according to claim 16 for treating and/or preventing alopecia, and/or for stimulating hair growth, and/or for growing hair.

20. The method of using the external preparation according to claim 16 for treating and/or preventing rhinitis.

21. (canceled)

22. (canceled)

23. The method according to claim 16, wherein X.sup.1-X.sup.12 are selected from the group consisting of following (1)-(12): (1) X.sup.1 denotes Gly, (2) X.sup.2 denotes Arg, Gln or His, (3) X.sup.3 denotes Lys or Arg, (4) X.sup.4 denotes Met, Leu or Ile, (5) X.sup.5 denotes Ile, (6) X.sup.6 denotes Ser or Gly, (7) X.sup.7 denotes Ser or Ala, (8) X.sup.8 denotes Ser, Gln, Val, Ala, Thr, Leu, Ile or Met, (9) X.sup.9 denotes Ser, Val, Ala or Thr, (10) X.sup.10 denotes Gly or Arg, (11) X.sup.11 denotes Leu, and (12) X.sup.12 denotes Gly, or a derivative thereof or a pharmaceutically acceptable salt thereof.

24. The method according to claim 16 selected from the cyclic peptides of SEQ ID NOs: 3-8 and SEQ ID NOs: 16-75, or a derivative thereof or a pharmaceutically acceptable salt thereof.

25. The method according to claim 16, wherein the derivative is substituted by a substituent which is capable of replacing a hydrogen atom, hydroxyl group, carboxy group, amino group or imino group in the cyclic peptide.

26. The method according to claim 16, wherein the cyclic peptide is formed by deleting 1 to 4 amino acids in the cyclic peptide expressed by any one of the Formulae (I-a)-(I-e), or by replacing them with or adding them other amino acids, and wherein the cyclic peptide has an equal function with the cyclic peptide expressed by each of said formulae.

27. The method according to claim 16, wherein the external preparation is an ingredient for a dermatitis therapeutic, dermatitis prophylactic, antipruritic, antiphlogistic, wound epithelialization-accelerating agent or skin-care product.

28. The method according to claim 27, wherein the skin-care product is used for moisturizing, and/or for preventing or improving rough skin, and/or for sebum/acne care, and/or for alleviating irritation/anti-inflammation, and/or for skin-lightening, and/or for anti-aging, and/or for preventing/alleviating ultraviolet lesion, and/or for slimming, and/or for skin-cleansing.

29. The method according to claim 16, wherein the external preparation is a bath agent, a body-cleansing agent or a hair-cleansing agent.

30. The method according to claim 16, wherein the external preparation is an alopecia therapeutic, an alopecia prophylactic, a hair growing agent and/or a hair growth stimulant.

31. The method according to claim 16, wherein the external preparation is a rhinitis therapeutic and/or rhinitis prophylactic.

32. The method according to claim 16, wherein the external preparation is a cosmetic.

33. The method according to claim 16, wherein the external preparation is a solid, semi-solid, powder, liquid, spray, ointment, cream, emulsion, gel or patch formulation.

34. The method according to claim 16, wherein the external preparation is used as a pharmaceutical product, a quasi-drug or a cosmetic product.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0038] FIG. 1

[0039] A diagram showing the result of applying BNP cyclic peptide (A) or a gel formulation (B) on either left or right side of the face of a subject having large wrinkles on the face.

[0040] FIG. 2

[0041] A diagram showing the effect before and after the application of either B ring gel formulation or BNP gel formulation to a patient with female pattern alopecia and alopecia pityroides.

MODE FOR CARRYING OUT THE INVENTION

[0042] Hereinafter, the present invention is explained in detail based on its suitable embodiments.

1. The cyclic peptide, a derivative thereof and a pharmaceutically acceptable salt thereof Firstly, the cyclic peptide of the invention, a derivative thereof and a pharmaceutically acceptable salt thereof are explained.

[0043] The cyclic peptide of the present invention has an amino acid sequence expressed by Formula I:

##STR00003##

wherein,
X.sup.1 denotes Gly, Val, Ala, Ser or Thr,
X.sup.2 denotes Arg, Gln or His,
X.sup.3 denotes Lys or Arg,
X.sup.4 denotes Met, Leu or Ile,
X.sup.5 denotes Ile or Val,
X.sup.6 denotes Ser or Gly,
X.sup.7 denotes Ser or Ala,
X.sup.8 denotes Ser, Gln, Val, Ala, Thr, Leu, Ile or Met,
X.sup.9 denotes Ser, Val, Ala or Thr,
X.sup.10 denotes Gly or Arg,
X.sup.11 denotes Leu, Met, Ile, Val or Ala,
X.sup.12 denotes Gly, Ser or Ala,
the line connecting two Cys denotes a disulfide bond,
and wherein the amino acid sequence does not have a peptide bond that is not between the amino acids constituting the amino acid sequence.

[0044] Such cyclic peptide is, similar to previously known BNPs, considered to bind to a receptor NPR-A (also known as GC-A) having a guanylate cyclase domain and promote the production of cyclic guanosine monophosphate (cGMP), and has activities such as, for example, diuretic action, vasodilation, renin-aldosterone secretion suppressing action, sympatholytic activity and hypertrophy suppressing action. It has a superior drug efficacy and effect, particularly an excellent immediate effect, as compared to BNP.

[0045] In addition, as described hereinbelow, the above cyclic peptide can be used as an ingredient of an external preparation for a dermatitis therapeutic/prophylactic, a rhinitis therapeutic/prophylactic, an alopecia therapeutic/prophylactic, a hair growing agent, a hair growth stimulant, an antipruritic, etc., or as an ingredient of a skin-care product, a quasi-drug or a cosmetic product. The above cyclic peptide can also be used as an alternative for BNP in an medicament utilizing the activity of BNP as above, e.g., in an medicament for hypertension, unstable angina, acute myocardial infarction, edematous diseases, renal failure, cardiac failure, immune diseases, obesity or metabolic syndrome.

[0046] In another embodiment of the present invention, in the cyclic peptide of the invention, X.sup.1-X.sup.12 in the Formula (I) may be defined as one or more selected from the group consisting of following (1)-(12):

(1) X.sup.1 denotes Gly, Val, Ala, Ser or Thr,
(2) X.sup.2 denotes Arg, Gln or His,
(3) X.sup.3 denotes Lys or Arg,
(4) X.sup.4 denotes Met, Leu or Ile,
(5) X.sup.5 denotes Ile or Val,
(6) X.sup.6 denotes Ser or Gly,
(7) X.sup.7 denotes Ser or Ala,
(8) X.sup.8 denotes Ser, Gln, Val, Ala, Thr, Leu, Ile or Met,
(9) X.sup.9 denotes Ser, Val, Ala or Thr,
(10) X.sup.10 denotes Gly or Arg,
(11) X.sup.11 denotes Leu, Met, Ile, Val or Ala, and
(12) X.sup.12 denotes Gly, Ser or Ala.

[0047] A preferred cyclic peptide of the present invention is a cyclic peptide of Formula (I), wherein X.sup.2 denotes Arg, X.sup.4 denotes Met, X.sup.6 denotes Ser, X.sup.7 denotes Ser, and/or X.sup.10 denotes Gly (SEQ ID NO: 2).

[0048] Furthermore, in another embodiment of the invention, in the cyclic peptide of the invention, the amino acid sequence expressed by Formula (I) may be selected from SEQ ID NOs: 3-8 and SEQ ID NOs: 16-75.

[0049] Furthermore, in the cyclic peptide of the invention, the amino acid sequence expressed by Formula (I) is preferably selected from the amino acid sequences expressed by the Formula (I-a)-Formula (I-e):

##STR00004##

wherein, the line connecting two Cys denotes a disulfide bond.

[0050] Each of the amino acid sequences expressed by these Formulae (I-a)-(I-e) is the cyclic part of human BNP (Formula (I-a)), swine BNP (Formula (I-b)), rat BNP (Formula (I-c)), rabbit BNP (Formula (I-d)) and murine BNP (Formula (I-e)), respectively. Therefore, the cyclic peptide having such an amino acid sequence will exhibit the aforementioned effects with more certainty. Because swine BNP cyclic peptide is consistent with those of avian (SEQ ID NO: 8, Cys-Phe-Gly-Arg-Arg-Ile-Asp-Arg-Ile-Gly-Ser-Leu-Ser-Gly-Met-Gly-Cys, wherein, 1st Cys and 17th Cys form a disulfide bond), bovine (SEQ ID NO: 9, Cys-Phe-Gly-Arg-Arg-Leu-Asp-Arg-Ile-Gly-Ser-Leu-Ser-Gly-Leu-Gly-Cys, wherein, 1st Cys and 17th Cys form a disulfide bond), feline (SEQ ID NO: 10, Cys-Phe-Gly-Arg-Arg-Leu-Asp-Arg-Ile-Gly-Ser-Leu-Ser-Gly-Leu-Gly-Cys, wherein, 1st Cys and 17th Cys form a disulfide bond), canine (SEQ ID NO: 11, Cys-Phe-Gly-Arg-Arg-Leu-Asp-Arg-Ile-Gly-Ser-Leu-Ser-Gly-Leu-Gly-Cys, wherein, 1st Cys and 17th Cys form a disulfide bond), and ovine (SEQ ID NO: 12, Cys-Phe-Gly-Arg-Arg-Leu-Asp-Arg-Ile-Gly-Ser-Leu-Ser-Gly-Leu-Gly-Cys, wherein, 1st Cys and 17th Cys form a disulfide bond), BNP cyclic peptides derived from these organisms also have similar effects as the cyclic peptide of the present invention. Among those mentioned above, it is particularly preferred that the amino acid sequence expressed by Formula (I) is the amino acid sequence expressed by Formula (I-a). In another embodiment of the invention, the cyclic peptide may be expressed by SEQ ID NOs: 16-75.

[0051] The cyclic peptide of the present invention encompasses a derivatized form of said cyclic peptide. Such a derivative can be used as such, as an active substance, or also used as a prodrug.

[0052] A derivative of the present invention can be obtained by adding (modifying) a known substituent to certain group of an amino acid in the cyclic peptide such as, for example, a hydrogen atom, hydroxyl group, carboxy group, amino group or imino group, or replacing it with a known replaceable substituent. Modification include, but not limited to, chemical modifications such as, for example, glycosylation, acetylation, phosphorylation and lipidation, to an amino acid within the cyclic peptide.

[0053] The addition (modification) or replacement of an amino acid within the cyclic peptide can occur in one group, or can occur in more than one group at the same time. Any known substituents can be used as long as being capable of replacing above groups and it goes without saying that such substituents naturally include, for example, protecting groups such as BOC.

[0054] The cyclic peptide of the invention further includes a mutated form of said cyclic peptide. Namely, a mutant of the present invention can be obtained by deleting an amino acid in the cyclic peptide, replacing it with or adding it another amino acid. The number of amino acid to be deleted, replaced with other amino acids or added is 4 or less, more preferably 3 or less, even more preferably 2 or less, and particularly preferable 1 or less. It also goes without saying that the deletion, replacement or addition of amino acids in the cyclic peptide may occur concurrently and independently to each other.

[0055] An amino acid that is capable of being replaced with another amino acid can be, in the case of human BNP (Formula (I-a)), exemplified as follows, without being limited thereto. The third amino acid from left, Gly, may be replaced with either Val, Ala, Ser or Thr. The forth amino acid from left, Arg, may be replaced with either Gln or His. The fifth amino acid from left, Lys, may be replaced with Arg. The sixth amino acid from left, amino acid, Met, may be replaced with either Leu or Ile. The ninth amino acid from left, Ile, may be replaced with Val. The twelfth amino acid from left, Ser, may be replaced with either Gln, Val, Ala, Thr, Leu, Ile or Met. The thirteenth amino acid from left, Ser, may be replaced with either Val, Ala or Thr. The fourteenth amino acid from left, Gly, may be replaced with Arg. The fifteenth amino acid from left, Leu, may be replaced with either Met, Ile, Val or Ala. The sixteenth amino acid from left, Gly, may be replaced with either Ser or Ala. Table 32 summarizes examples of replaceable amino acids in the cyclic peptide of the invention, though the invention will not be limited thereto.

[0056] Cyclic peptides in which one amino acid has been replaced include such as, for example, SEQ ID NOs: 16-44, though the invention will not be limited thereto. Cyclic peptides in which two amino acids have been replaced include such as, for example, SEQ ID NO: 45-58, though the invention will not be limited thereto. Yet cyclic peptides in which three amino acids have been replaced include such as, for example, SEQ ID NOs: 59-70, though the invention will not be limited thereto. Furthermore, cyclic peptides in which four amino acids have been replaced include such as, for example, SEQ ID NOs: 71-75, though the invention will not be limited thereto. It also goes without saying that five or more amino acids can be replaced by appropriately combining the aforementioned amino acid.

[0057] Deletion of one to several amino acids can take place in similar manner to the replacement of the aforementioned amino acids with other amino acids.

[0058] The number of amino acid which are to be deleted or replaced with other amino acids or added may be determined such that the cyclic peptide will have 80% homology, preferably 90% homology to the cyclic peptide of the invention.

[0059] In addition, the present invention may be a mutant as described above, and a derivative thereof. As long as it retains the effect of the invention, any mutant or a derivative thereof is encompassed in the cyclic peptide according to the invention. In another embodiment, it has at least a BNP activity. For purpose of improving the activity of the cyclic peptide of the invention, prolonging the effect of the invention, and/or increasing storage stability of the cyclic peptide of the invention, the cyclic peptide of the invention or the amino acids constituting said peptide may be altered in an appropriate manner. For example, an amino acid in the cyclic peptide of the invention may be chemically modified, some amino acids constituting the cyclic peptide may be deleted or replaced with other amino acids, and/or new amino acids may be added.

[0060] For example, the C-terminal group COOH of one Cys of the cyclic peptide may be replaced with COOR.sup.1, CONHR.sup.1 or CONR.sup.12, and/or the N-terminal group NH.sub.2 of the other Cys of the cyclic peptide may be replaced with NHC(O)R.sup.1 or N(C(O)R.sup.1).sub.2. Here, each appearance of R.sup.1 is independently a branched or straight hydrocarbon group or an alkylene glycol chain or sugar chain having 1 to 20 carbon atoms. The number of carbon atoms in R.sup.1 is preferably 1 to 10, more preferably 1 to 5, yet more preferably 1 to 2.

[0061] Pharmaceutically acceptable salts include, without being particularly limited, as the cyclic peptide of the invention or a derivative, such as for example, a salt with an inorganic base, a salt formed with an organic base, a salt formed with an inorganic acid, a salt formed with an organic acid, a salt formed with a basic or acidic amino acid. Examples of suitable salt formed with inorganic base include such as, for example, an alkaline metal salt such as a sodium salt and potassium salt; an alkaline earth meal salt such as a calcium salt and magnesium salt; and an aluminum salt and ammonium salt. Examples of suitable salt formed with organic base include such as, for example, a salt with an alkyl amine such as trimethyl amine or triethyl amine; a salt formed with a heterocyclic amine such as pyridine and picoline; a salt formed with an alkanol amine such as ethanol amine, diethanol amine and triethanol amine; a salt formed with a cycloalkyl amine such as cyclohexyl amine and dicyclohexyl amine; a salt formed with an alkylene diamine derivative such as N,N-dibenzylethylenediamine. Examples of suitable salt formed with inorganic acid include such as, for example, a salt formed with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid or phosphoric acid. Examples of suitable salt formed with organic acid include such as, for example, a salt formed with a monocarboxylic acid such as formic acid, acetic acid, trifluoroacetic acid and propionic acid; a salt formed with a polyvalent carboxylic acid such as fumaric acid, oxalic acid and maleic acid; a salt formed with a oxycarboxylic acid such as tartaric acid, citric acid, succinic acid and malic acid; a salt formed with a sulfonic acid such as methane sulfonic acid, benzene sulfonic acid and p-toluene sulfonic acid; and a salt formed with benzoic acid. Examples of suitable salt formed with a neutral amino acid include such as, for example, a salt formed with glycine, valine or leucine; examples of suitable salt formed with a basic amino acid include such as, for example, a salt formed with arginine, lysine or ornithine; and examples of suitable salt formed with an acidic amino acid include such as, for example, a salt formed with aspartic acid acid or glutamic acid.

[0062] Among those mentioned above, a cyclic peptide composed of the amino acid sequence expressed by Formula (I) or a pharmaceutically acceptable salt is preferred. Namely, it is preferred that the amino acid sequence expressed by Formula (I) is not replaced.

[0063] A method for producing the cyclic peptide of the present invention, a derivative and pharmaceutically acceptable salt thereof may employ, without being particularly limited, for example, any known chemical synthetic or genetic engineering methods.

[0064] When the amino acid sequence as above is chemically synthesized, it may be synthesized by any chemical synthetic method, or any known method for peptide synthesis, for example, or solid-phase or liquid-phase synthetic method. Moreover, any commercial synthesizer (e.g., SHIMADZU Corporation: PSSM-8) may be used for synthesis.

[0065] A disulfide bond can be formed in the amino acid sequence, for example, by DMSO oxidation method or iodine oxidation method without being particularly limited. In this case, an intramolecular disulfide bond can be formed by treating a free sulfhydryl group or a sulfhydryl group protected by a protecting group with either DMSO or iodine (I2) to result in said cyclic peptide.

[0066] A protecting group includes such as, for example, 4-methylbenzyl group (Bzl (4Me)), trityl group (Trt), tert-butyl group, N-(acetyl) aminomethyl group (Acm). Deprotection can be carried out through appropriate treatment corresponding to these protecting group, for example, for 4-methylbenzyl group by treating with a strong acid, and for N-(acetyl) aminomethyl group by treating with iodine.

[0067] Next, if necessary, the cyclic peptide is derivatized to result in a derivative. Derivatization can be carried out by known method. Alternatively, a derivative of the cyclic peptide can be produced at the time of peptide synthesis by introducing in advance a substituent into the amino acid constituting the cyclic peptide.

[0068] Then, if necessary, a pharmaceutically acceptable salt is produced by ion exchange of the cyclic peptide or derivative thereof. Ion exchange can be carried out, for example, by bringing into the contact the cyclic peptide or derivative thereof with a desired acid or base.

[0069] On the other hand, when the peptide is synthesized by genetic engineering, it can be synthesized by any known method such as, for example, methods described in Sambrook J. et al., Molecular Cloning, A Laboratory Manual (4th edition) (Cold Spring Harbor Laboratory Press (2012)). For instance, a DNA fragment coding for the amino acid sequence expressed by Formula (I) is prepared at first. Preparation of the DNA fragment can be performed, for example, if it is a case in which the DNA fragment codes for the amino acid sequence expressed by Formula (I-a), by amplifying the DNA fragment by PCR using a vector comprising a full-length human BNP gene as template and primers designed to synthesize a defined DNA region. Alternatively, a DNA fragment can be chemically synthesized.

[0070] Then, the amplified DNA fragment is ligated into an appropriate vector to obtain a recombinant vector for protein expression. Next, the vector for protein expression is allowed to be taken up by target cells and the transformed cells are selected. Finally, the protein produced by the cells (i.e., the protein composed of the amino acid sequence expressed by Formula (I)) is collected.

[0071] Formation of disulfide bond, derivatization and salt formation in the collected protein can be carried out as described above.

[0072] The identification of the compound of interest such as a cyclic peptide can be confirmed by known procedures such as, for example, reverse-phase HPLC or mass spectroscopy.

[0073] The presence or absence of BNP activity in the obtained compound can readily be confirmed by known means. For instance, it can be confirmed by examining cGMP producing activity in NPR-A receptor-expressing cells.

2. External Preparation

[0074] Secondly, the external preparation of the present invention is explained.

[0075] The external preparation of the present invention comprises one or more cyclic peptide and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof as described above. If two or more cyclic peptides are used, the number of different cyclic peptide to be mixed is not limited, though 2 to 3 are preferred in terms of preparation cost and convenience. It includes SEQ ID NO: 19, 29, 33, 36, 39 and 43, for example, but the invention is not limited to these and these cyclic peptide can appropriately be combined as long as it does not interfere the effect of the invention.

2.1 Application

[0076] The external preparation of the present invention can be used for following uses, for example, without being particularly limited, and in each case exerts remarkable effect which had not previously been achieved. The external preparation of the present invention is not limited to any particular use, and can be used as a pharmaceutical product, a quasi-drug and/or a cosmetic product depending on its drug efficacy and effect.

[0077] Hereinbelow, the invention will be described in detail for each of its use.

(1) Therapeutic/Prophylactic for Dermatitis

[0078] The external preparation of the present invention can be a a therapeutic and/or prophylactic for dermatitis.

[0079] The external preparation of the present invention can, upon being applied onto the subject's skin or mucosa suffering dermatitis, quickly relieve or eliminate various perceptible symptoms and conditions that are or can be caused by dermatitis such as pruritus, soreness (pain), hot sensation, tautness, erythema, infiltration, papule, lichenification, crust, exudation or skin desiccation, improving the symptoms of dermatitis. In addition, the external preparation of the present invention does not cause any irritation at the site of application.

[0080] Furthermore, such action of the external preparation comprising the cyclic peptide of the invention is effected more quickly and for prolonged time period as compared to that of an external preparation comprising BNP having the tail part. Although reasons for these advantageous effects of the cyclic peptide over such BNP are not clear, the cyclic peptide of the invention is considered to have an advantageous structure for binding to its receptor (NPR-A receptor) over BNP, thereby enabling faster binding to the NPR-A receptor in the vicinity of the affected site and at the same time prolonging its binding time. Moreover, it is considered that its relatively quick binding to the NPR-A receptor near the affected site can prevents the cyclic peptide from diffusing out of the affected site via bloodstream, etc., enabling it to stay around the affected site for relatively long time.

[0081] The external preparation of the present invention does not only suppress or prevent inflammation in dermatitis but also acts to restore and/or retain the barrier function of skin. Here, skin's barrier function works for protecting skin against entry of stimuli and saprophytes from external environment or for retaining moisture. The external preparation restores the barrier function, and thus able to prevent the progress or exacerbation of inflammation. The barrier function of skin is greatly affected by the alignment state of corneocytes in the corneum, i.e., state of skin texture and moisturization. The external preparation of the present invention acts to improve skin texture and to moisturize skin. Therefore, its effects of restoring and maintaining skin barrier function are also obvious.

[0082] The external preparation of the present invention is also effective on skin symptoms commonly called acne, i.e., such as comedones, red papule, pustula, cysts/tuberosity.

[0083] Moreover, a quick relief or elimination of perceptible symptoms or conditions will reduce the burden of the subject (patient) and improve the QOL of the patient, while preventing the patient from being bothered by itch or soreness and from acting to damage the affected site, for example, touching or scratching. This effect of the external preparation of the present invention to relieve or eliminate perceptible symptoms or conditions is, in general, exhibited within 10 minutes, preferably within 5 minutes, more preferably within 3 minutes after application.

[0084] In addition, the external preparation of the present invention can treat dermatitis in which steroid dermatosis has been developed, or dermatitis that is intractable by other common drugs for dermatitis therapy such as steroid and tacrolimus, for example dermatitis for which a sufficient therapeutic effect cannot be achieved by these formulations, or dermatitis which is resistant to these formulations, or dermatitis for which the use of these formulations is not suitable or desirable. Conventional widely-used steroid external preparations have significant problems upon terminating application that the severity returns to the pre-application level and that a rebound phenomenon may occur and exacerbate the condition. The external preparation of the present invention has no such problems as a rebound phenomenon.

[0085] Dermatitis is in general a disease which causes inflammation in skin or mucosa. Dermatitis is normally accompanied with one or more symptoms selected from acute eczema symptoms such as erythema, infiltrative erythema, papule, vesicle, pustula, infiltration, incrustation and desquamation; chronic eczema symptoms such as lichenification and pigmentation; and scales, crust (scab) attachment, scratching, scratch scar, prurigo nodularis, erosion, edema, oozing and squamatization.

[0086] Dermatitis herein is not particularly limited as long as it is a disease which is accompanied with inflammation in skin or mucosa and includes such as, for example, eczema and atopic dermatitis such as chronic eczema, dyshidrotic eczema, infantile xerotic eczema; contact dermatitis such as allergic contact dermatitis and primary irritant contact dermatitis; seborrheic dermatitis; asteatotic dermatitis; autosensitization dermatitis; stasis dermatitis; urticaria such as allergic urticaria (e.g., alimentary urticaria and drug-induced urticaria) and nonallergic urticaria (e.g., physical urticaria, solar urticaria and cholinergic urticaria); insect bite; drug eruption; psoriasis such as plaque psoriasis, guttate psoriasis, erythrodermic psoriasis, pustular psoriasis and psoriasis arthropathica; prurigo such as chronic prurigo, acute prurigo, gestational prurigo and nodular prurigo; rosacea; rosacea-like dermatitis; cutaneous vasculitis such as cutaneous allergic vasculitis; cutaneous pruritus such as systemic cutaneous pruritus, localized cutaneous pruritus, senile cutaneous pruritus and gestational pruritus; solar dermatitis; erythrosis; nummular dermatitis; localized scratching dermatitis; perioral dermatitis; pompholyx; keratosis pilaris; lichen planus, dyshidrotic eczema, dyshidrosis, miliaria and acne vulgarisacne vulgaris. The external preparation of the present invention can be applied to the therapy and prophylaxis of any of these diseases.

[0087] The external preparation of the present invention exerts an excellent effect especially on eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, insect bite, allergic or nonallergic urticaria and psoriasis, preferably on eczema, atopic dermatitis, contact dermatitis and insect bite, more preferably on eczema and atopic dermatitis. Therefore, the external preparation of the present invention can be used for the purpose of the therapy and prophylaxis of at least one or more dermatitis selected from above-mentioned group.

(2) Ingredients of Cosmetic or Skin-Care Product

[0088] The external preparation of the present invention may be an ingredient for a cosmetic or skin-care product.

[0089] The external preparation of the present invention will, upon being applied to skin and/or mucosa, provide moisture to the applied site and exert moisturizing effect, preventing skin from dryness and roughness. In cases where corneum is present at the site of application, it exerts an effect of improving skin texture there. It also provides and retains moderate elasticity and flexibility of skin and mucosa, softens skin, gives skin and mucosa resilience and firmness. Moreover, it improves wrinkles (including fine and large wrinkles) and flabbiness at the applied site, and further diminishes the appearance of dullness and spots. By diminishing dullness and spots away, it consequently provides a skin-lightening effect.

[0090] As a result, the external preparation of the present invention can be used for the purpose of maintaining or improving skin and/or mucosal condition, without being particularly limited, for example, for obtaining at least one effects selected from a group consisting of preventing or improving dry skin, skin roughness, sensitive skin, roughened lips and appearance of large and fine wrinkles, maintaining skin or mucosal condition, anti-aging, and skin-lightening, or obtaining for at least one effects selected from the above effects. Skin roughness herein includes miliaria, chilblain, cracked skin, chapped skin, acne, diaper rashes, festering, chafed inner thighs and razor burns.

[0091] Specific utilities of the external preparation of the present invention as an ingredient for the cosmetic or skin-care product includes, without being particularly limited, for example, cosmetics for make-up such as face lotion, emulsion, serum, cream, cold cream, gel, mask, pack, powder, hand soap, perfume, deodorant, as well as foundation, face powder, eye shadow, eyeliner, mascara, eyebrow, blush, makeup base, lip stick, lip cream and nail polish, and furthermore cosmetics for hair such as shampoo, rinse, conditioner, hair color, hair tonic, styling agent and perm chemical, body cleansers such as face wash, cleansing and body soap, skin-care products such as body powder, after-shave lotion and pre-shave lotion, and bath agents.

[0092] The above effects of the external preparation of the present invention are quickly exhibited, within 10 minutes in general, preferably within 5 minutes, more preferably within 3 minutes after application depending on the type of the effect.

[0093] The above effects of the external preparation of the present invention last for relatively long time. In general, the effect lasts for 4 hours or more, preferably, 8 hours or more, more preferably 24 hours or more after it started to effect depending on the type of the effect.

(3) Antipruritic

[0094] The external preparation of the present invention can be an antipruritic.

[0095] As mentioned above, the external preparation of the present invention is suitable for using as antipruritic because it quickly exerts an excellent antipruritic effect at the applied site when being applied to skin or mucosa.

[0096] The antipruritic effect of the external preparation of the present invention as mentioned above is exhibited within 10 minutes in general, preferably within 5 minutes, more preferably within 3 minutes after application.

(4) Alopecia Therapeutic, Alopecia Prophylactic, Hair Growing Agent and Hair Growth Stimulant

[0097] The external preparation of the present invention can also be an alopecia therapeutic, alopecia prophylactic, hair growing agent and/or hair growth stimulant.

[0098] When the external preparation of the present invention is applied to a site of hair loss or a site where hair growth is to be stimulated, it acts to prevent hair loss and promote hair growth stimulation and hair growing at the applied site. It has effects such as nourishing hair, promoting hair growth, and improving or preventing hair thinning at the applied site. In this case, hairs to be stimulated to grow tend to become terminal hairs or hairs which are not white hairs. These effects are exhibited in relatively early stage as compared to other active ingredients previously used in alopecia therapeutics e.g., BNP, and the effects obtained are significant.

[0099] The external preparation of the present invention also has an improving or preventing effect on dermatitis as mentioned above. Therefore, it can improve or prevent skin inflammation associated with alopecia. Such effects are particularly advantageous when alopecia has been exacerbated due to skin condition of the site of application (such as scalp).

[0100] Moreover, the external preparation of the present invention exerts moisturizing and skin texture-improving effects at the applied site when being applied to skin as mentioned above. It can remove and suppress dandruff and itching, while giving moisture to hair and scalp, improving and preventing dryness and keeping hair and scalp healthy. It also can improve seborrhea.

[0101] The external preparation of the present invention also acts to alleviate or eliminate the perceptible symptoms or conditions as mentioned above, thereby reducing the burden of the subject (patient) and improving QOL of the patient. Moreover, it can prevent the patient from being bothered by itch or soreness and acting to damage the affected site, for example, touching or scratching, thereby preventing exacerbation of skin condition which may induce alopecia.

[0102] When the external preparation of the present invention is used as alopecia therapeutic or prophylactic, applicable alopecia includes such as, for example, alopecia as listed below, without being particularly limited. The external preparation can be used for the purpose of treating or preventing one ore more of these alopecia.

(Acquired Alopecia)

[0103] (i) Alopecia without accompanying scarring or skin lesion (alopecia areata, male pattern alopecia, seborrheic alopecia, alopecia pityroides, female pattern alopecia, gestational alopecia, malignant alopecia, senile alopecia, alopecia totalis, alopecia areata multilocularis, ophiasis, drug-induced alopecia, cancer chemotherapy-induced alopecia and radiation exposure-induced alopecia, traumatic/mechanical alopecia, malnutrition/metabolic disorder associated alopecia, endocrine dysfunction associated alopecia and telogen effluvium (post partum alopecia, alopecia after high fever)).
(ii) Alopecia observed on skin lesion or pathologic skin (infection-induced alopecia, tumor-induced alopecia, inflammation-induced alopecia)
(iii) Scarring alopecia (skin infection-induced alopecia, alopecia induced by infiltration of inflammatory cells)

(Congenital Alopecia)

[0104] Diffuse alopecia, congenital atrichia, alopecia in hereditary syndromes, localized alopecia, phakomatosis, aplasia cutis, congenital alopecia triangularis.

[0105] Among those mentioned above, the external preparation of the present invention exerts an excellent effect particularly on acquired alopecia, preferably on alopecia without accompanying scarring or skin lesion, more preferably on alopecia areata, male pattern alopecia, seborrheic alopecia, alopecia pityroides, female pattern alopecia, gestational alopecia, malignant alopecia, senile alopecia, alopecia totalis, alopecia areata multilocularis, ophiasis, drug-induced alopecia, cancer chemotherapy-induced alopecia and radiation exposure-induced alopecia, traumatic/mechanical alopecia, malnutrition/metabolic disorder associated alopecia, endocrine dysfunction associated alopecia and telogen effluvium. Therefore, the external preparation of the present invention can be used for the purpose of treating or preventing at least one or more alopecia selected from the above-mentioned group.

(5) Rhinitis Therapeutic and/or Prophylactic

[0106] The external preparation of the present invention can also be a rhinitis therapeutic and/or prophylactic.

[0107] Rhinitis is a disease caused by mucosal inflammation in nasal cavity and/or paranasal cavity and induces main symptoms such as nasal obstruction, rhinorrhea, sudden recurrent sneezing, as well as symptoms such as pruritus. In the present invention, rhinitis does not only includes rhinitis in a narrow definition which are accompanied with mucosal inflammation in nasal cavity, but also includes sinusitis accompanied with mucosal inflammation in paranasal cavity.

[0108] The external preparation of the present invention can improve or prevent various symptoms associated with rhinitis such as nasal obstruction, rhinorrhea, sneezing and pruritus, when being applied to nasal cavity mucosa and/or paranasal cavity mucosa. Especially these effects are exhibited quickly and last for a prolonged time as compared to a rhinitis therapeutic comprising BNP having the tail part.

[0109] Such effects of the external preparation of the present invention on rhinitis are exhibited within 8 minutes in general, preferably within 5 minutes, more preferably within 3 minutes after its application.

[0110] Also the effect of the external preparation of the present invention on rhinitis as described above lasts for 4 hours or more in general, preferably for 8 hours or more, more preferably for 24 hours or more after it started to effect.

[0111] When the external preparation of the present invention is used as a rhinitis therapeutic and/or prophylactic, applicable rhinitis includes such as, without being particularly limited, for example, infectious rhinitis including acute rhinitis and chronic rhinitis; hypersensitive non-infectious rhinitis including combined (flower hypersensitivity) rhinitis, rhinitis with rhinorrhea, congestive rhinitis, edematous rhinitis and dry-nose type rhinitis; irritant-induced rhinitis including physical rhinitis, chemical rhinitis and radiation rhinitis; and atrophic rhinitis and idiopathic granulomatous rhinitis; and sinusitis such as acute sinusitis, chronic sinusitis (maxillary empyema), eosinophilic sinusitis and paranasal cavity mycosis, The external preparation can be used for the purpose of treating or preventing one ore more of the above.

[0112] Combined rhinitis includes, for example, allergic rhinitis including perennial allergic rhinitis and seasonal allergic rhinitis, and nonallergic rhinitis including vasomotor (essential) rhinitis and eosinophilic rhinitis.

[0113] Rhinitis with rhinorrhea includes, for example, gustatory rhinitis, cold air-induced rhinitis and senile rhinitis.

[0114] Congestive rhinitis includes, for example, drug-induced rhinitis, psychogenic rhinitis, gestational rhinitis, endocrine rhinitis and cold rhinitis.

[0115] Edematous rhinitis includes, for example, aspirin-hypersensitive rhinitis.

[0116] Among those mentioned above, the external preparation of the present invention exerts an excellent effect particularly on hypersensitive non-infectious rhinitis, irritant-induced rhinitis and sinusitis, preferably on hypersensitive non-infectious rhinitis and chronic sinusitis, more preferably on combined (flower hypersensitivity) rhinitis, rhinitis with rhinorrhea, congestive rhinitis, edematous rhinitis and dry-nose type rhinitis, chronic sinusitis. Therefore, the external preparation of the present invention can be used for the purpose of treating or preventing at least one or more rhinitis selected from the above-mentioned group.

[0117] In terms of symptoms, since the external preparation of the present invention acts as described above, it can be used for any of rhinitis with sneezing/rhinorrhea, rhinitis with nasal obstruction and rhinitis with both symptoms.

[0118] In addition, the external preparation of the present invention exhibits an effect of improving rhinitis which cannot be cured by a rhinitis therapeutic that has conventionally been used such as steroid drug.

(6) Other External Preparations

[0119] The external preparation of the present invention can also be used for an application other than those described above for the effect of the cyclic peptide as described above. In this case, the external preparation of the present invention can set an objective that is not the effects of the cyclic peptide as described above. In this case, the cyclic peptide of the invention, a derivative and/or a pharmaceutically acceptable salt thereof is used for the purpose of assisting the main effect of the external preparation or adding another effect to the main effect.

[0120] Such application include such as, without being particularly limited, for example, body powder, deodorant, depilation agent, soap, body shampoo, bath agent, hand soap, perfume, sunscreen, and antiinflammatory agent and antifungal agent.

2.2 Formulation

[0121] The external preparation of the present invention can exert the effect of its active ingredient, i.e., the cyclic peptide or a derivative and/or pharmaceutically acceptable salt thereof, certainly and quickly in the vicinity of the applied site by being locally applied to the site of the interest (e.g., affected site) on skin or mucosa.

[0122] Such external preparation can be, without being particularly limited, for example, an agent for integument, eye drop, ear drop, nasal drop, buccal agent or suppository. Among these, when the external preparation of the present invention is a dermatitis therapeutic, a dermatitis prophylactic, an antipruritic, a skin-care product, an alopecia therapeutic, an alopecia prophylactic, a hair growing agent or a hair growth stimulant, it is preferably an agent for integument. On the other hand, when the external preparation of the present invention is a rhinitis therapeutic and/or prophylactic, it is preferably a nasal drop. Furthermore, when it is a therapeutic/prophylactic of a corneal disease, it is preferably an eye drop.

[0123] When the external preparation of the present invention is an agent for integument, it can be, without being particularly limited, for example, an external solid formulation, an external liquid formulation, a spray formulation, an ointment, an emulsion, a cream, a gel formulation or a patch.

[0124] An external solid formulation is a solid formulation for applying or spraying onto such as skin. Such an external solid formulation includes, for example, a powder form external formulation.

[0125] An external liquid formulation is a liquid formulation for applying onto such as skin. Such an external liquid formulation includes, for example, a lotion and liniment.

[0126] A spray formulation is a formulation for spraying an active ingredient in a mist, powder, foam or paste form onto skin. Such spray formulation includes, for example, an external aerosol and a pump spray formulation.

[0127] An ointment is a semi-solid formulation which is applied to skin and comprises an active ingredient dissolved or dispersed in a base. The ointment can also be a lip cream for locally applying to lips, etc.

[0128] A cream is a semi-solid formulation which is applied to skin and emulsified as either oil-in-water or water-in-oil type.

[0129] A gel formulation is a gelled formulation which is applied to skin. The gel formulation includes, for example, aqueous gel formulation and oil-based gel formulation.

[0130] A patch is a formulation which is attached to skin. The patch includes, for example, a tape or plaster.

[0131] A nasal drop is a formulation which is administered to nasal cavity or nasal mucosa. The nasal drop includes, for example, nasal powder formulation and a nasal drop. Among these, nasal drop is preferred.

[0132] In any formulation described above, the external preparation of the present invention comprises the cyclic peptide of the present invention and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof as described above.

[0133] When the formulation is an external liquid formulation, an ointment, a cream or a gel formulation, the external preparation of the present invention comprises the cyclic peptide and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof at a concentration of, for example, 0.0001 to 1000000 g/g, preferably 0.001 to 10000 g/g, more preferably 0.01 to 1000 g/g, yet more preferably 0.1 to 100 g/g. In another embodiment, it may comprises the cyclic peptide at a concentration of 1 to 800 g/g or 3 to 500 g/g.

[0134] When the formulation is a spray formulation, the external preparation of the present invention comprises in the stock solution of the spray formulation the cyclic peptide and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof at a concentration of, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, 0.01 to 1000 g/mL, more preferably 0.1 to 100 g/mL, yet more preferably 1 to 100 g/mL. In another embodiment, it may comprises the cyclic peptide at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

[0135] When the formulation is a patch, the external preparation of the present invention comprises the cyclic peptide and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof at a concentration of, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, it may comprises the cyclic peptide at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

[0136] When the formulation is a nasal drop, especially a liquid nasal drop, the external preparation of the present invention comprises in the nasal drop solution (nasal drop) the cyclic peptide and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof at a concentration of, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, it may comprises the cyclic peptide at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

[0137] The external preparation of the present invention can be formulated, for any formulation as described above, by using methods and constituent materials known to those skilled in the art.

[0138] Available constituent materials include such as, without being particularly limited, for example, a gelator, oily ingredient, higher alcohol, fatty acid, ultraviolet absorbing agent, ultraviolet scattering agent, powder, pigment, surfactant, polyhydric alcohol/sugar, polymer, bioactive ingredient, solvent, antioxidant, flavor and antiseptic agent.

[0139] Various organic or inorganic gelator compounds can be used.

[0140] An inorganic gelator compound includes such as, for example, hydrous or water-absorbable silicate, for example, aluminum silicate (e.g., bentonite), magnesium-aluminum silicate and colloidal silica.

[0141] As an organic gelator compound, a natural, semisynthetic or synthetic polymer can be used. natural and semisynthetic polymers include such as, for example, polysaccharides such as cellulose, starch, tragacanth, gum arabic, xanthane gum, agar, gelatin, alginic acid and a salt thereof (e.g., sodium alginate and a derivative thereof), lower alkylcellulose (e.g., methylcellulose or ethylcellulose), carboxy- or hydroxy-lower alkylcellulose (e.g., carboxymethylcellulose or hydroxypropylcellulose). A synthetic polymer includes such as, for example, carboxylvinyl polymer, sodium polyacrylate, (vinylmethyl ether/ethyl maleate) copolymer, polymethacrylate, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylate or polymethacrylate. Alternatively, as a gel formulation, commercially available gelators such as, for example, Lubrajel NP, Lubrajel CG, Lubrajel DV, Lubrajel MS, Lubrajel OIL, Lubrajel TW, Lubrajel DS (Ashland Inc.) can also be used.

[0142] As an oily ingredient, for example, various ester, ether, hydrocarbon, silicone and fluorine oil phase ingredient, as well as animal and plant oils and a hardened oil thereof, and waxes of natural origin.

[0143] The ester oil phase ingredients include such as, for example, glyceryl tri(2-ethyl hexanoate), cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, butyl myristate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldocecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate, isoalkyl neopentanoate, glyceryl tri(capryl-caprinate), trimethylol propane tri(2-ethylhexanoate), trimethylol propane triisostearate, pentaerythritol tetra(2-ethylhexaonate), cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isocetyl myristate, isostearyl myristate, isocetyl palmitate, isostearyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, cetostearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprinate, propylene glycol di(capryl-caprinate), propylene glycol dicaprylate, neopentyl glycol dicaprinate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecanoate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerol oleic acid ester, polyglycerol isostearic acid ester, dipropyl carbonate, dialkyl (C12-18) carbonate, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di(2-ethylhexyl) succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate and isostearyl 12-stearoylhydroxystearate.

[0144] Hydrocarbon oil phase ingredients include such as, for example, squalane, liquid paraffin, -olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, solid paraffin, polybutene, microcrystalline wax and vaseline.

[0145] Silicone oil phase ingredients include such as, for example, dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, dacamethylpolysiloxane, dodecamethylcyclosiloxane, methyl hydrogen polysiloxane, polyether-denatured organo(polysiloxane), dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane/methylstearoxysiloxane copolymer, alkyl-denatured organo(polysiloxane), terminal-denatured organo(polysiloxane), amino-denatured silicone oil, amino-denatured organo(polysiloxane), dimethiconol, silicone gel, acryl silicone, trimethyl siloxysilicate, silicone RTV gum.

[0146] Fluorine oil phase ingredients include such as, for example, perfluoropolyether, fluorine-denatured organo(polysiloxane), pitch fluoride, fluorocarbon, fluoroalcohol and fluoroalkyl-polyoxyalkylene co-denatured organo(polysiloxane).

[0147] Animal and plant oils and a hardened oil thereof, and waxes of natural origin include such as, for example, animal and plant oils and the hardened oil thereof such as beef tallow, hardened beef tallow, lard, hardened lard, horse oil, mink oil, orange roughy oil, fish oil, hardened fish oil, egg yolk, jojoba oil; plant oils and a hardened oil thereof such as avocado oil, almond oil, olive oil, cacao butter, apricot kernel oil, kukui nut oil, sesame oil, wheat germ oil, rice germ oil, rice bran oil, safflower oil, shea butter, soybean oil, evening primrose oil, camellia oil, corn oil, rapeseed oil, hardened rapeseed oil, palm kernel oil, hardened palm kernel oil, palm oil, hardened palm oil, peanut oil, hardened peanut oil, castor oil, hardened castor oil, sunflower oil, grape seed oil, jojoba oil, hardened jojoba oil, macadamia nut oil, meadowfoam oil, cottonseed oil, hardened cottonseed oil, palm oil, hardened palm oil, rose hip oil; waxes such as beeswax, beeswax having high acid value, lanolin, reduced lanolin, hardened lanolin, liquid lanolin, carnauba wax and montan wax.

[0148] Higher alcohols include such as, for example, lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, behenyl alcohol, 2-ethyl hexanol, hexadecyl alcohol, octyl dodecanol.

[0149] Fatty acids include such as, for example, caprylic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachic acid, arachidonic acid, behenic acid, erucic acid, 2-ethyl hexanoic acid.

[0150] Ultraviolet absorbing agents include such as, for example, paraminobenzoate, amyl paraminobenzoate, ethyl dihydroxypropyl paraminobenzoate, glyceryl paraminobenzoate, ethyl paraminobenzoate, octyl paraminobenzoate, octyl dimethyl paraminobenzoate, ethylene glycol salicylate, octyl salicylate, triethanol aminesalicylate, phenyl salicylate, butyl phenyl salicylate, benzyl salicylate, menthyl salicylate, benzyl cinnamate, octyl paramethoxy cinnamate, 2-ethylhexyl paramethoxy cinnamate, glyceryl diparamethoxy cinnamate mono(2-ethylhexanoate), isopropyl paramethoxy cinnamate, paramethoxyhydrocinnamate diethanol amine salt, diisopropyl-disopropyl cinnamate ester mixture, urocanate, urocanate ethyl, hydroxymethoxy benzophenone, hydroxymethoxy benzophenone sulfonate and a salt thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone sulfonate, dihydroxybenzophenone, dihydroxy dimethoxybenzophenone, hydroxyoctoxybenzophenone, tetrahydroxybenzophenone, butyl methoxy-dibenzoyl methan, 2,4,6-trianilino-p-(carbo-2-ethylhexyl-1-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl) benzotriazole, methyl-O-aminobenzoate, 2-ethylhexyl-2-cyano-3,3-diphenyl acrylate, phenylbenzimidazole sulfate, 3-(4-methyl benzylidene) camphor, isopropyl dibenzoylmethan, 4-(3,4-dimethoxyphenylmethylene)-2,5-dioxo-1-imidazolidine propionate 2-ethylhexyl, and polymer derivatives or silane derivative thereof, titanium oxide and zinc oxide and dispersion thereof. The zinc oxide and titanium oxide may be surface-treated.

[0151] Dermal absorption auxiliary agents include such as, for example, acetic acid, sodium acetate, limonene, menthol, salicylic acid, hyaluronic acid, oleic acid, N,N-diethyl-m-toluamide (N,N-diethyl-3-methyl benzamide), n-butyl stearate, benzyl alcohol, isopropyl myristate, isopropyl palmitate, polypropylene glycol, crotamiton, diethyl sebacate, N-methylpyrrolidone, N-ethylpyrrolidone and lauryl alcohol.

[0152] Powders and pigments include such as, for example, pigments such as Pigment Red 104, Pigment Red 201, Pigment Yellow 4, Pigment Blue 1, Pigment Black 401, basic dyes, HC colors, disperse dyes, direct colors, lake pigments such as Pigment Yellow 4 AL lake, Pigment Yellow 203 BA lake; polymers such as nylon powder, silk powder, urethane powder, silicone powder, methyl polymethacrylate powder, cellulose powder, starch, silicone elastomer spherical powder and polyethylene powder; colored pigments such as yellow iron oxide, red iron oxide, black iron oxide, chromic oxide, carbon black, ultramarine, iron blue; white pigments such zinc oxide, titanium oxide, cerium oxide; extender pigment such as tare, mica, sericite, kaolin and tabular barium sulfate; pearl pigment such as titanium mica; metal salt such as barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate and magnesium silicate; inorganic powder such as silica, alumina; metal soap such as aluminum stearate, magnesium stearate, zinc palmitate, zinc myristate, magnesium myristate, zinc laurate, zinc undecylenate; bentonite, smectite, boron nitride, etc. The shape (spherical, rod-like, needle-like, tabular, amorphous, scaly, spindle-shaped) and particle diameter of these powders are not particularly limited.

[0153] These powders and pigments may be pre-treated by known conventional surface processing such as, for example, fluorine compound processing, silicone processing, silicone resin processing, pendant processing, processing with silane coupling agent, processing with titanium coupling agent, oil solution processing, N-acylated lysine processing, polyacrylic acid processing, metal soap processing, amino acid processing, lecithin processing, inorganic compound processing, plasma processing and mechanochemical processing.

[0154] As the surfactant, any of an anionic surfactant, a cationic surfactant, ampholytic surfactant and non-ionic surfactant can be used as appropriate.

[0155] Anionic surfactants include such as, for example, fatty acid soap, -acylsulfonate, alkylsulfonate, alkylallylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkylethersulfate, alkylamidesulfate, alkylphosphate, POE alkylphosphate, alkylamidephosphate, alkyloylalkyl taurine salt, N-acylamino acid salt, POE alkylether carboxylate, alkylsulfosuccinate, sodium alkylsulfoacetate, acylisethionate, acylated hydrolyzed collagen peptide salt and perfluoroalkyl phosphate ester.

[0156] Cationic surfactants include such as, for example, alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, propyldimethylhydroxypropylammonium behenic amide chloride, diethylaminoethyl-stearamide, dimethylaminopropyl-stearamide and lanolin derivative quaternary ammonium salt. Cationic surfactants also include tertiary amines such as fatty acid amide dialkyl amine and salts thereof.

[0157] Ampholytic surfactants include various ampholytic surfactants, for example, those of carboxybetain type, amidebetain type, sulfobetain type, hydroxysulfobetain type, amidesulfobetain type, phoshobetain type, aminocarboxylate type, imidazoline derivative type and amideamine type.

[0158] Non-ionic surfactants include such as, for example, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE sorbitol fatty acid ester, POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POE-POP copolymer, POE-POP alkyl ether, polyether-denatured silicone alkanolamide laurate, alkylamine oxide, hydrogenated soybean phospholipid, hydrogenized soybean phospholipid, polymer surfactant and biosurfactant.

[0159] Natural surfactants may also be used, including such as, for example, lecithin, saponin and saccharide surfactant.

[0160] Polyhydric alcohols and sugars include such as, for example, ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, diglycerin, polyglycerin, 3-methyl-1,3-butanediol, 1,3-butylene glycol, sorbitol, mannitol, raffinose, erythritol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, trehalose, alkylated trehalose, mixed isomerized sugar, sulfated trehalose and pullulan. Chemically modificated forms of these can also be used.

[0161] Polymers include such as, for example, anionic polymer compounds such as acrylate ester/methacrylate ester copolymer, vinyl acetate/crotonate copolymer, vinyl acetate/crotonate/vinyl neodecanoate copolymer, methylvinyl ether maleate half ester, t-butyl acrylate/ethyl acrylate/methacrylate copolymer, vinylpyrrolidone/vinyl acetate/vinyl propionate copolymer, vinyl acetate/crotonatecopolymer (RUBISET CA: BASF), vinyl acetate/crotonate/vinylpyrrolidone copolymer, vinylpyrrolidone/acrylate copolymer, acrylate/acrylamide copolymer, vinyl acetate/butyl maleate/isoisobornyl acrylate copolymer, carboxyvinyl polymer, acrylate/methacrylate alkyl copolymer, and ampholytic acetate of dialkyl aminoethyl methacrylate polymer, ampholytic polymer compound such as octyl acrylamide acrylate/hydroxypropyl acrylate/butyl methacrylateaminoethyl copolymer, quaternized compound of vinylpyrrolidone/dimethylaminoethyl methacrylate, cationic polymer compounds such as methylvinyl imidazolium chloride/vinylpyrrolidone copolymer, non-ionic polymer compounds suchs as polyvinylpyrrolidone, vinylpyrrolidone/vinyl acetate copolymer, vinylpyrrolidone/dimethylaminoethyl methacrylate copolymer, vinylcaprolactam/vinylpyrrolidone/dimethylaminoethyl methacrylate copolymer. Polymer compounds of natural origin such as cellulose or derivatives thereof, keratin and collagen or derivatives thereof, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, high-methoxyl pectin, low methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabino galactan, can be karaya, gum tragacanth, alginates, albumin, casein, curdlan, gellan gum and dextran, and glucooligosaccharide, fucose containing polysaccharide, rhamnose containing polysaccharide can also suitably be combined.

[0162] Bioactive ingredients include substances which provide skin with some bioactivity when being applied to skin. Bioactive ingredients include those with such as, for example, skin-lightening, anti-inflammatory, anti-aging, ultraviolet protection, slimming, firming, antioxidation, hair growth stimulating/hair growing, suppressing hair growth, moisturizing, promoting circulation, antimicrobial/sterilization, cool/warm feeling, promoting wound cure, alleviating irritation, analgesic and cell-activating effects. Bioactive ingredients include such as plant extracts, seaweed extracts, vitamins and derivatives thereof, amino acids, various peptides other than the cyclic peptide, biopolymers such as sodium hyaluronate and mucopolysaccharide, intercellular lipid constituents such as ceramide, phytosphingosine, cholesterol and phytosterol, and analogues thereof, and enzymatic ingredients.

[0163] Examples of suitable combining ingredient include such as, for example, Angelica keiskei extract, avocado extract, sweet Hydrangea leaf extract, althea extract, arnica extract, aloe extract, apricot extract, apricot kernel extract, isoflavones, ginkgo extract, fennel extract, turmeric extract, oolong tea extract, Rose Fruit extract, echinacea extract, Baikal skullcup extract, Amur Corktree extract, Coptis japonica extract, barley extract, Hypericum erectum extract, Lamium album extract, cress extract, orange extract, cacao extract, desiccated sea water, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, pumpkin seed extract, chamomile extract, carrot extract, Artemisia capillaris extract, licorice extract, hibiscus extract, Pyracantha fortuneana extract, kiwi extract, cinchona extract, cucumber extract, guanosine, gardenia extract, Sasa veitchii extract, Sophora flavescens extract, cranberry extract, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentian extract, tea extract, yeast extract, burdock extract, rice bran fermentation extract, rice germ oil, comfrey extract, collagen, cowberry extract, Asarum sieboldii Miq. extract, bupleurum extract, umbilical cord extract, salvia extract, soapwort extract, bamboo grass extract, crataegus extract, zanthoxylum extract, shiitake mushroom extract, Rehmannia root extract, Lithospermum root extract, Perilla frutescens extract, Tilia japonica (Miq.) extract, Filipendula multijuga extract, Paeonia lactillora extract, calamus extract, white birch extract, Equisetum arvense extract, Hedera helix extract, Crataegus oxyacantha extract, European elder extract, yarrow extract, peppermint extract, sage extract, tree mallow extract, cnidium extract, extract of Swertia japonica (Schult.) Makino, soybean extract, jujube extract, soybean fermentation extract, thyme extract, tea extract, clove extract, cogon extract, orange peel extract, Oenothera tetraptera extract, Centella asiatica extract, Terminalia sericea extract, dong quai extract, common marigold extract, peach kernel extract, bitter orange peel extract, extract of Houttuynia cordata Thunb., tomato extract, natto extract, ginseng extract, garlic extract, wild rose extract, hibiscus extract, Ophiopogon japonicus extract, parsley extract, honey, banana flower extract, hamamelis extract, parietaria extract, Isodon japonicus (Burm.) Hara extract, bisabolol, loquat extract, coltsfoot extract, butterbur flower extract, Poria Sclerotium extract, butcher's-broom extract, grape extract, propolis, luffa extract, safflower extract, peppermint extract, linden extract, tree peonyextract, hop extract, pine extract, horse chestnut extract, Lysichiton camtschatcense (L.) Schott extract, Sapindus mukurossi Gaertn. extract, Melissa extract, peach extract, bluebottle extract, eucalyptus extract, Saxifraga stolonifera extract, Citrus junos extract, coix seed extract, mugwort extract, lavender extract, apple extract, lytchee (Litchi) extract, lettuce extract, lemon extract, Chinese milk vetch extract, rose extract, rosemary extract, Roman chamomile extract and royal jelly extract.

[0164] Examples also include such as biopolymers such as deoxyribonucleic acid, mucopolysaccharides, sodium hyaluronate, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan and hydrolyzed eggshell membrane; moisturizing ingredients such as amino acids, hydrolyzed peptides, sodium lactate, urea, sodium carbonate pyrrolidone, betain, whey, trimethyl glycine, polypeptides such as lysine/arginine condensate etc.; intercellular lipid constituents such as sphingolipid, ceramide, phytosphingosine, cholesterol, cholesterol derivative, phytosterol derivative, phospholipid and analogues thereof; anti-inflammatory agents such as -aminoaminocaproic acid, glycyrrhizic acid, -glycyrrhetinic acid, lysozyme chloride, guaiazulen, hydrocortisone, tea tree oil, vitamins such as vitamin A and derivatives thereof, vitamin B2 and derivatives thereof, vitamin B6 and derivatives thereof, vitamin C and derivatives thereof, vitamin D and derivatives thereof, vitamin E and derivatives thereof, calcium pantothenate, biotin and nicotinic acid; active agents such as allantoin, diisopropylamine dichloroacetate, 4-aminomethylcyclohexane carbonate; antioxidants such as tocopherol, carotenoids, flavonoids, tannin, lignan and saponin; cell activating agents such as -hydroxy acid and -hydroxy acid; circulation accelerating agents such as -oryzanol and vitamin E derivative; wound curing agents such as retinol and retinol derivative; skin-lightening agents such as arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linolenic acid, tranexamic acid, glutathione; hair growing agents such as cepharanthine, glycyrrhiza extract, capsicum tincture, hinokitiol, garlic extract iodide, pyridoxine hydrochloride, DL--tocopherol, DL--tocopherol acetate, nicotinic ac id, nicotinic acid derivative, calcium pantothenate, D-pantothenyl alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropylmethylphenol, estradiol, ethinylestradiol, carpronium chloride, Benzalkonium chloride, diphenhydramine hydrochloride, Takanal, camphor, salicylic acid, vanillyl nonylate amide, vanillyl nonanoate amide, piroctone olamine, glyceryl pentadecanoate, L-menthol, mononitroguaiacol, resorcin, -aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharides tincture, Cyclosporine, zinc pyrithione, hydrocortisone, minoxidil, polyoxyethylene sorbitan monostearate, peppermint oil, Sasanishiki extract.

[0165] Antioxidants include such as, for example, sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tolyl biguanide, nordihydroguaiaretinoic acid, parahydroxyanisole, butyl hydroxyanisole, dibutyl hydroxytoluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoids, flavonoids, tannin, lignans, saponins and plant extracts in which the antioxidative effect is recognized such as apple extract or clove extract.

[0166] Solvents include such as physiological saline, purified water, ethanol, lower alcohols, ethers, LPG, fluorocarbons, N-methylpyrrolidone, fluoroalcohols, volatile straight chain silicones and next-generation chlorofluorocarbons.

[0167] The cyclic peptide of the invention and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof can be used for preparation of the external preparation as described above, and therefore the present invention also relates to a use of the cyclic peptide of the invention and/or a derivative thereof and/or a pharmaceutically acceptable salt thereof for preparing the external preparation.

3. Method of Using the External Preparation

[0168] Next, methods of using the external preparation of the invention is explained.

[0169] A method of using the external preparation of the invention comprises applying the external preparation of the present invention as described above to the skin and/or mucosa of a subject.

[0170] Subjects include human and vertebrates such as, without being particularly limited, for example, birds and mammals. Animals specifically include, for example, experimental animals including rodents such as mice, rats, gerbils, hamsters and guinea pigs, domestic animals such as pigs, goats, horses, sheep and minks, pet animals such as dogs and cats, primates such as human, monkeys, cynomolgus monkeys, rhesus monkeys, marmosets, orangutans and chimpanzees. On the other hand, human may be excluded from the subjects.

[0171] Skin and/or mucosa of a subject to which the external preparation is to be applied may be skin or mucosa at any site of the subject, and the external preparation is applicable, for example, to skin or mucosa of head (scalp), face, neck, arms, torso, arms, hands or feet, etc.

[0172] Specific method for using the external preparation of the present invention is as follows.

(1) Method of Treating and Preventing Dermatitis

[0173] When the external preparation of the present invention is used as a therapeutic/prophylactic of dermatitis, the external preparation can be applied directly to the aimed site of skin and/or mucosa (e.g., the affected site where dermatitis has been developed).

[0174] The application frequency is, without being particularly limited, for example, 1 to 10 times a day, preferably 1 to 5 times a day, yet more preferably 1 to 3 times a day. Because the external preparation of the present invention has a long lasting time, it exerts a sufficient effect even if it is applied at relatively low frequency, for example, once a day.

[0175] In terms of dosage, the total amount of the cyclic peptide of the invention and a derivative thereof and a pharmaceutically acceptable salt thereof for one application can be, without being particularly limited, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, each dose may comprise the cyclic peptide of the invention at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

(2) Method of Alleviating or Resolving Itch

[0176] When the external preparation of the present invention is used as an antipruritic, the external preparation can be applied directly to the aimed site of skin and/or mucosa.

[0177] The application frequency is, without being particularly limited, for example, 1 to 10 times a day, preferably 1 to 5 times a day, yet more preferably 1 to 3 times a day.

[0178] In terms of dosage, the total amount of the cyclic peptide of the invention and a derivative thereof and a pharmaceutically acceptable salt thereof for one application can be, without being particularly limited, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, each dose may comprise the cyclic peptide of the invention at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

(3) Method of Using as a Cosmetic

[0179] When the external preparation of the present invention is used as a cosmetic, the external preparation can be applied directly to the aimed site of skin and/or mucosa.

[0180] The application frequency is, without being particularly limited, for example, 1 to 10 times a day, preferably 1 to 5 times a day, yet more preferably 1 to 3 times a day.

[0181] In terms of dosage, the total amount of the cyclic peptide of the invention and a derivative thereof and a pharmaceutically acceptable salt thereof for one application can be, without being particularly limited, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, each dose may comprise the cyclic peptide of the invention at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

(4) Method of Treating and Preventing Alopecia, and Method of Stimulating Hair Growth and Method of Growing Hair

[0182] When the external preparation of the present invention is used as a therapeutic/prophylactic of alopecia, a hair growth stimulant or a hair growing agent, the external preparation can be applied directly to the aimed site (e.g., decalvant site of scalp or skin)

[0183] The application frequency is, without being particularly limited, for example, 1 to 10 times a day, preferably 1 to 5 times a day, yet more preferably 1 to 3 times a day.

[0184] In terms of dosage, the total amount of the cyclic peptide of the invention and a derivative thereof and a pharmaceutically acceptable salt thereof for one application can be, without being particularly limited, for example, 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, each dose may comprise the cyclic peptide of the invention at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

(5) Method of Treating and Preventing Rhinitis

[0185] When the external preparation of the present invention is used as a therapeutic/prophylactic of rhinitis, the external preparation can be applied directly to the aimed site (e.g., nasal cavity mucosa).

[0186] The application frequency is, without being particularly limited, for example, 1 to 10 times a day, preferably 1 to 5 times a day, yet more preferably 1 to 3 times a day.

[0187] In terms of dosage, without being particularly limited, for example, for one application to each nasal cavity, the total amount of the cyclic peptide of the invention and a derivative thereof and a pharmaceutically acceptable salt thereof can be 0.0001 to 1000000 g/mL, preferably 0.001 to 10000 g/mL, more preferably 0.01 to 1000 g/mL, yet more preferably 0.1 to 100 g/mL, particularly preferably 1 to 100 g/mL. In another embodiment, each dose may comprise the cyclic peptide of the invention at a concentration of 1 to 800 g/mL or 3 to 500 g/mL.

4. Medicament

[0188] Next, the medicament of the present invention is explained.

[0189] The medicament of the present invention comprises one or more cyclic peptides of the invention or derivatives thereof or pharmaceutically acceptable salts thereof.

[0190] As mentioned above, similarly to BNP, the cyclic peptide of the present invention or a derivative thereof or a pharmaceutically acceptable salt thereof binds to the receptor NPR-A (also known as GC-A), which has a guanylate cyclase domain, and promotes the production of cyclic guanosine monophosphate (cGMP). It is considered to have effects such as, for example, diuretic action, vasodilation, renin-aldosterone secretion suppressing action, sympatholytic activity and hypertrophy suppressing action. Also, the compounds of the invention is considered to have a superior efficacy and effect, particularly a superior immediate effect, as compared with BNP.

[0191] Therefore, the medicament of the present invention can be used for a similar object as a conventional medicament comprising BNP as an active agent.

[0192] Diseases for which the medicament of the present invention is applicable include such as, without being particularly limited, for example, the aforementioned various diseases as well as hypertension, unstable angina, acute myocardial infarction, edematous diseases, renal failure, cardiac failure, immune diseases, obesity and metabolic syndrome.

[0193] A formulation of the medicament of the present invention can be, without being particularly limited, addingly to the external preparations as mentioned above, for example, formulations for oral administration such as tablets, capsules, granules, powders, oral solution, syrup and oral jelly, formulations for oral application such as buccal tablets, buccal spray formulation, buccal semi-solid formulation and mouthwash, formulations for administration via injection such as an injection and infusion, dialysis formulation such as dialysis solution, as well as an inhalant, eye drop, ocular ointment, ear drop, vaginal tablet and vaginal suppository.

Working Examples

[0194] Hereinbelow, the present invention is further specifically illustrated by working examples. It should be noted that the present invention is not limited by these working examples.

1. Preparation of the Cyclic Peptide

[0195] Firstly, a cyclic peptide composed of the amino acid sequence expressed by Formula I-a is synthesized.

[0196] Specifically, a linear peptide consisting of 17 amino acids was formed by sequentially binding amino acids by solid-phase peptide synthesis using a peptide synthesizer. Subsequently, protecting groups at Cys1 and Cys17 were detached before treating with iodine (I2) to form a cysteine binding between oxidatively same amino acid residues, thereby forming a cyclic peptide.

[0197] A composition comprising the obtained cyclic peptide was purified by reverse-phase high performance liquid chromatography (reverse-phase HPLC) and then lyophilized to yield a purified cyclic peptide as white powder.

[0198] Mass spectroscopy was performed on the obtained cyclic peptide.

[0199] Conditions for HPLC are shown below:

Apparatus: Agilent 1100

[0200] Flow rate: 1.0 ml/min
Eluent A: 0.1% trifluoroacetic acid/water
Eluent B: 0.1% trifluoroacetic acid/acetonitrile
Gradient: 80% Eluent B, isocratic

[0201] Conditions for Mass spectroscopy (MS) are shown below:

Apparatus: Thermo Finnigan LCQ Advantage

[0202] Ionization method: electrospray ionization
Analytical method: ion trapping

[0203] The observed results, m/z=901.83 ([M+2H].sup.2+), m/z=1801.84 ([M+H].sup.+), confirmed that above peptide is in agreement with the theoretical values of the molecular weight (1802.07) and the mass number (1800.8069) calculated from the composition formula of the cyclic peptide of interest (C.sub.72H.sub.120N.sub.24O.sub.24S.sub.3).

[0204] Furthermore, the purity of the above peptide was measured by HPLC using following conditions:

[0205] Column: Discovery C18, 4.6 mm250 mm, particle diameter 5 micron

[0206] Column temperature: room temperature

[0207] Eluent A: 0.1% trifluoroacetic acid/water

[0208] Eluent B: 0.1% trifluoroacetic acid/acetonitrile

[0209] Gradient: 10 to 30% Eluent B/20 minutes

[0210] Flow rate: 1.2 ml/min

[0211] Temperature: room temperature

[0212] Injected volume: 20 l

[0213] Detector: UV detector (detection wavelength: 215 nm)

[0214] The result of measurement confirmed that the purity of the obtained protein was 99.2%.

2. Preparation of Formulation

2.1 Preparation of Gel Formulation

[0215] Preparation of a gel-based formulation comprising a cyclic peptide composed of the amino acid sequence expressed by Formula I-a (in the working examples below, B ring is referred to mean a cyclic peptide expressed by such Formula I-a) (B ring gel formulation, Working Examples) and a gel-based formulation comprising human BNP (BNP gel formulation, Comparative Examples) were carried out as follows.

[0216] 0.1 g of methyl-p-hydroxybenzoate (trade name: Mekkins-M, Ueno Fine Chemicals Industry, Ltd.), 0.2 g of phenoxyethanol and 3.0 g of 1,2-pentanediol were weighed into one same vessel, heated to 60 to 70 C. to make a homogenous solution, and this solution was poured into a mixer.

[0217] Next, added into the mixer 6.0 g of concentrated glycerin, and then the mixture of 0.44 g of carboxyvinyl polymer (trade name: Carbopol 940, Lubrizol Advanced Materials Corporation) and 0.08 g of xanthane gum (trade name: KELTROL T, CP Kelco, Inc.), and the mixuter was stirred with a paddle until they dispersed sufficiently.

[0218] Then 83.95 g of purified water was gradually added with stirring with a paddle. The mixer was heated to 70 to 80 C. while stirring with a paddle or disper until the dispersed contents were dissolved to give a solution. Subsequently, the disper was stopped and the solution was cooled immediately after confirming that the contents in the solution were dissolved. When the temperature of the mixer reached approximately 40 C., 6.0 g of Lubrajel NP from Ashland Inc. (glycerin 2.7 g, carboxyvinyl polymer 0.06 g, sodium polyacrylate 0.018 g, water 3.222 g) was added to the solution, mixed uniformly with a paddle. Subsequently, 0.230 g of potassium hydroxide was further added to neutralize the solution, then the rotation of the paddle was stopped when the temperature of the mixer reached 25 C. to prepare a gel base.

[0219] Next, 20.1 mg of the the cyclic peptide (B ring) composed of the amino acid sequence expressed by Formula I-a was dissolved in 144 mL of physiological saline to obtain B ring solution. 0.131 mL of this solution was admixed with 10 g of the gel base obtained as described above, and the mixture was stirred uniformly to prepare a gel based formulation (B ring gel formulation) containing B ring at a concentration of about 1 M (about 1.8 g/g). Similarly, gel based formulations containing B ring at concentrations of about 0.3 M (about 0.54 g/g), about 0.5 M (about 0.9 g/g) and about 2.0 M (about 3.6 g/g) were prepared. In the working examples below, as long as it is specifically described otherwise, B ring gel formulation used contained B ring at a concentration of about 1 M.

[0220] Next, 20.5 mg of human BNP-32 (American Peptide Company) was dissolved in 118 mL of physiological saline to obtain BNP solution. 0.2 mL of this solution was admixed with 10 g of the gel base obtained as described above, and the mixture was stirred uniformly to prepare a gel based formulation (BNP gel formulation) containing BNP-32 at a concentration of about 1 M. Similarly, gel based formulations containing BNP-32 at concentrations of about 0.5 M and 2.0 M were prepared. In the working examples below, as long as being specifically described otherwise, BNP gel formulation used contained BNP at a concentration of about 1 M.

2.2 Preparation of Nasal Drop

[0221] A nasal drop comprising the cyclic peptide (B ring) composed of the amino acid sequence expressed by Formula I-a (B ring nasal drop, Working Examples) and a nasal drop comprising human BNP (BNP nasal drop, Comparative Examples) were prepared as follows.

[0222] Firstly, the cyclic peptide (B ring) composed of the amino acid sequence expressed by Formula I-a was dissolved in physiological saline, the concentration was adjusted to prepare B ring nasal drop containing B ring at a concentration of about 1 mol/1 (about 1.8 g/g).

[0223] Next, B ring nasal drop was filled in a quantitative nasal spray container (AS ONE Corporation) and the amount to be sprayed for each administration was adjusted to 100 l (0.1 ml), resulting B ring nasal drop.

[0224] Similarly, a BNP nasal drop comprising human BNP (American Peptide Company) at a concentration of 1 mol/1 was obtained, filled in a quantitative nasal spray container (AS ONE Corporation), and the amount to be sprayed for each administration was adjusted to 100 l (0.1 ml), resulting BNP nasal drop.

3. Confirming Therapeutic Effect on Dermatitis

3.1 Confirming the Effect of B Ring Gel Formulation

[0225] For subjects suffering various dermatitis, B ring gel formulation as described was applied onto the affected site, and changes in symptoms before and after the application were observed. When possible, as a comparative example, BNP gel formulation as described was applied onto another affected site on the same subject where B ring gel formulation had not been applied, and changes in symptoms before and after the application were observed. For pruritus, VAS (Visual Analogue Scale) was used for evaluating each affected site on 10 stages.

[0226] The examination results are shown in Tables 1 to 7 along with age, sexuality, symptoms of the subject, and formulation given to the subject.

TABLE-US-00001 TABLE 1 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D1 2 M atopic Face: presented with Face: aches removed 2 min after 1 face 8.fwdarw.0 Age: 30's B ring gel dermatitis erythema, scales and skin application. Desiccation, scales and right arm 7.fwdarw.0 Sex: male formulation desiccation (moderate erythema After 3 days of 1 application disease). application, erythema, scales, Limbs: presented with and desiccation remarkably remitted erythema, lichenification, (insignificant). skin desiccation and Right forearm: itches stopped to some scratch scar (severe). extent after 1.5 min on B ring-applied site. No itching or tingling after 2 min. remitteed after 4 min. Desiccation improved. Skin moisturized and soft, with natural appearance. No subjective discomfort (moderate). 2 M Face: realized after 4 min. face 8.fwdarw.4 BNP gel erythema and scales more distinguished left arm 7.fwdarw.4 formulation than B ring side. 1 application Left forearm: after 4 min, skin was still dry. No improvement in lichenification. Rough and stiff skin (severe). D2 1 M atopic Flushing, infiltrating erythema, Right back: itches stopped after 3 min. 8.fwdarw.0 Age: 20's B ring gel dermatitis scales, many scratch scars Flushing erythema, infiltration and Sex: male formulation erythroderma (severe) accompanied by scales remitted. Skin became soft 1 application strong itches causing sleep (moderate). 1 M disruption and desire to Left back: still iching after 3 min. 8.fwdarw.4 BNP gel scratch. no remission of flushing erythema, formulation infiltration and scales (severe). 1 application D3 0.5 M atopic Presented with infiltrating Right back: no tingling itches 30 sec 6.fwdarw.0 Age 10's B ring gel dermatitis erythema, scales, papules, after application. Skin felt normal. Sex: male formulation (moderate) accompanied by Erythema, infiltration and scales 1 application tingling itches. remitted after 4 min. (mild). 0.5 M Left back: still itching after 4 min, 6.fwdarw.6 BNP gel with no remission of erythema, formulation infiltration and scales (moderate). 1 application indicates data missing or illegible when filed

TABLE-US-00002 TABLE 2 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D4 2 M atopic Presented with infiltrating Right back: itches relieved just after 10.fwdarw.0 Age: 20's B ring gel dermatitis erythema, scales, many application. No itches after 2 min with Sex: male formulation scratch scars with exudation remission of erythema and infiltration 1 application (severe) accompanied by (moderate). strong itches that cause sleep Forehead: itches reduced after 1.5 min. disruption on the back, After 5 min skin was soft with no lichenification, erythema, lichenification. Erythema and scales scales and scratch scars on remitted. Wound quickly healed. Scratch forehead (severe). scars epithelized. 2 M Back: presented with many Left back: still itching after 4 min with no 10.fwdarw.3 BNP gel scratch scars with exudation, remission of erythema and infiltration formulation infiltration, erythema, (severe) 1 application crusts (severe). Forehead: no improvement in lichenification, desiccation and scales, with remaining itches (severe) D5 1 M atopic Presented with infiltrating Right face: itches, edema and infiltration 9.fwdarw.0 Age: 20's B ring gel dermatitis erythema, edema and crusts remitted/relieved just after application. Sex: male formulation (severe) accompanied by Skin felt no itch and moisturized after 1 1 application strong tingling ithces. min (moderate). Back: presented with many No itches at all after 2 min. Edema and scratch scars with exudation, erythema remitted. Right eye became infiltration, erythema, crusts easy to open. 1 M and papules (severe). Left face: still itching after 3 min with no 9.fwdarw.4 BNP gel remission of erythema, infiltration and formulation edema. Being difficult to open left eye 1 application (severe). D6 1 M atopic Presented with lichenification, Right back: itches improved/relieved 6.fwdarw.0 Age: 10's B ring gel dermatitis , erythema, after 3 min. Lichenification, Sex: male formulation scales, papules and many erythema, scales, scratch 1 application scratch scars (severe) scars all improved as compared to left. accompanied by strong itches. Skin was soft and moisturized Back: presented with many (moderate) 1 M scratch scars with exudation, Left back: still itching after 4 min with no 6.fwdarw.3 BNP gel infiltration, erythema, crusts remission of erythema and infiltration formulation and papules (severe). (severe) 1 application Forehead: lichenification, desiccation and scales not improved, with itches left (severe). indicates data missing or illegible when filed

TABLE-US-00003 TABLE 3 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D7 0.3 M atopic Presented with infiltrating No itches after 1 min. Erythema 10.fwdarw.0 Intractable Age: 20's B ring gel dermatitis erythema, papules, exudation remitted and exudation stopped stroid- Sex: male formulation and many scratch scars with after 10 min. 1 application/day resistance 1 application crust (severe) accompanied for 3 days cleared of infiltrating by strong itches that cause erythema, papules, exudation and sleep disruption. scratch scars. Remarkably improved crusts, partly left mild erythema (mild). D8 0.5 M atopic Presented with erythema, 5 days after application, erythema, 8.fwdarw.1 Intractable Sex: male B ring gel dermatitis infiltration, scales, infiltration, scales, crusts and steroid- formulation crusts, scratch scars and wound quickly cured with little resistance 1 application, skin desiccation (severe). scratch scars left (mild). 5 days Desiccation remarkably remitted Skin moisturized and soft. D9 B ring gel atopic Left forearm presented with Itches removed in 2 min (VAS 0). 10.fwdarw.0 Age: 30's formulation dermatitis many scratch scars with Erythema and exudation improved. Sex: male 2 applications/ exudation, infiltration, Severity of rash improved to day 1 day erythema, papules, moderate disease level after accompanied with tingling 5 min. After 1 more application strong itches (VAS10) given on the same night, (severe) there was no itches for 3 days thereafter. Scratch scars epithelized. D10 1 M atopic Presented with infiltrating Right forearm: itches topped after 8.fwdarw.0 Age: 50's B ring gel dermatitis erythema, scales, papules and 3 min. Erythema and infiltration Sex: female formulation many scratch scars (severe) improved (moderate). 1 application accompanied by intermediate 1 M strong itches Left forearm: still itching after 8.fwdarw.4 BNP gel 4 min with no remission of erythema formulation and infiltration (severe) 1 application D11 1 M atopic Presented with infiltrating Right face: immediately after 10.fwdarw.0 Age: 40's B ring gel dermatitis erythema, scales and many application, itches and skin tautness Sex: female formulation scratch scars (severe) relieved, with erythema getting better. 1 application accompanied by drastic No itching after 1 min. Erythema, itches and tautness. infiltration and desiccation Back: presented with many remitted (moderate). 1 M scratch scars with exudation, Left face: still itching after 3 min, 10.fwdarw.4 BNP gel infiltration, erythema, having erythema, infiltration and edema formulation crusts and papules (severe). with swollen feeling (severe). 1 application

TABLE-US-00004 TABLE 4 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D12 1 m atopic Strong tingling itches, Right face: itches remitted after 3 min. 10 .fwdarw. 1 Age: 40's B ring gel dermititis disrupted sleep, infiltrating Edema, erythema and infiltration started Sex: formulation erythema, scales, exudation, to be remitted. After 30 min, anti- female 1 application cracks, crusts, scratch scars; inflammatory effect observed, exudation eyes cannot be opened due to stopped, wound epithelization promoted, exudate sealing eyelids cracks almost epithelized. Edema, (severe). erythema and infiltration remitted to some extent. Eyelids became openable steadily. 1 m Left face: still having tingling itches after 10 .fwdarw. 6 BNP gel 3 min with no remission of erythema and formulation infiltration. Itches till left after 30 min. 1 application D13 1 m atopic Obvious infiltration, erythema Right face: erythema started to be 8 .fwdarw. 0 Age: 20's B ring gel dermititis and papules all over the face, remitted and itches removed just after (immediate Sex: formulation accompanied with strong application. Erythema and infiltration after female 1 application itches and hot flash (severe). remitted after 4 min (moderate). application) 1 m Left face: itching with hot flush and 8 .fwdarw. 0 BNP gel redness just after application. Itches (after 4 formulation removed after 4 min. minutes) 1 application D14 1 m atopic Hand eczema of which main Right hand: itches removed after 3 min 9 .fwdarw. 0 Steroid- Age: 20's B ring gel dermititis symptom is contact dermititis (VAS0). Lichenification improved, skin resistance; Sex: formulation due to detergent and got soft, erythema and infiltration intractable female 1 application disinfectant. Presented with remitted (moderate). lichenification 1 m lichenification, erythema, Left hand: no improvement in 9 .fwdarw. 4 which cannot BNP gel scales and scratch scars lichenification and erythema after 10 min be remitted formulation (severe). Sleep disruption by (severe). within this 1 application strong itches (VAS9). short time (3 min) by conventional therapy.

TABLE-US-00005 TABLE 5 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D15 1 m atopic Presented with erythema, Erythema, infiltration, scales and crusts 9 .fwdarw. 1 Age: 20's B ring gel dermititis infiltration, scales, crusts, wound epithelization improved 3 days Sex: formulation scratch scars (severe), with after application. Scratch scars female 1 application/ strong itches. significantly improved. Only mild day erythema and scales observed (mild). 3 days Desiccation relieved, improved and moisturzed skin texture. Itches and scratch scars significantly improved. D16 1 m contact Contact dermititis due to hair Right side: tingling itches removed after 8 .fwdarw. 0 Age: 60's B ring gel dermititis dye with erythena, edema 1 min. Itches eliminated after 3 min Sex: formulation and infiltration along hairline (VAS0), felt better than left. Erythema female 1 application (moderate disease) and infiltration remitted. accompanied by strong Anti-inflammatory effect observed. tingling itches (VAS8) Edema improved (mild). 1 m Left side: symptoms less improved than 8 .fwdarw. 4 BNP gel right (moderate). formulation 1 application D17 1 m chronic Presented with infiltrating Right face: itches stopped just after 10 .fwdarw. 0 Age: 40's B ring gel eczema erythema, scales, application. Erythema remitted. No Sex: formulation lichenification, crusts, papules scales and desiccation after 2 min. Skin female 1 application and skin desiccation (severe) was moisturized (moderate). accompanied by drastic itches Forehead: itches controlled after 1.5 min. and tautness. No lichenification after 5 min. Skin got soft. Erythema and scales remitted, scratch scars epithelized and wound quickly healed. 1 m Left face: still itching and taut after 4 min 10 .fwdarw. 5 BNP gel with no remission of desiccation, scales, formulation erythema and infiltration (severe). 1 application D18 1 m eczema Presented with erythema, After 15 min, erythema, edema, 8 .fwdarw. 0 Age: B ring gel edema, infiltration, scales and infiltration and scales significantly Sex: formulation papules (severe). remitted and skin texture improved female 1 application (mild). 1 m After 15 min, still many scales, 8 .fwdarw. 4 BNP gel erythema, edema and infiltration left. formulation Skin texture is rough and still itching. 1 application

TABLE-US-00006 TABLE 6 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D20 1 m wheal, Presented with multiple Right lower limb: itches removed just 10 .fwdarw. 0 Age: 20's B ring gel insect bites wheals, erythema and very after application. Erythema started to be Sex: formulation strong itches on both lower remitted after 1.5 min. Wheals female 1 application limbs. disappeared in about 1 hour. D31 1 m wheal, Presented with multiple Left lower limb: itches removed 1 min 10 .fwdarw. 0 Age: 30's B ring gel insect bites wheals and very strong itches after application, wheals remitted after 2 Sex: formulation on right lower limb. min. female 1 application D21 1 m childhood Presented with desiccation, Right back: itches stopped after 30 sec. 8 .fwdarw. 0 Age: 10's B ring gel xerotic many milary large papules, Desiccation, scales and erythema Sex: formulation eczema desquamation, strong pruritus, improved in a few minutes, Skin got female 2 applications/ and many scratch scars on moisturized. After 3 days, wounds day trunk; incrustation, erythema quickly epithelized leaving one scratch 3 days and lichenification due to scar. Skin is smooth and moisterized scratching (severe). with no desiccation (insignificant). 1 m Left back: desiccation, scales and 8 .fwdarw. 3 BNP gel erythema left after few minutes. formulation 1 application D32 1 m parapsoriasis Presented with multiple Scales, erythema and infiltration remitted N/A Age: 40's B ring gel guttata erythrokeratoderma with after 4 min. Sex: formulation sticking small scales on back. female 1 application D33 0.5 m psoriasis Both lower limbs presented Scales and erythema started to be N/A Age: 60's B ring gel vulgaris with many red plaques with remitted 10 min after 1st application. Sex: male formulation attached small scales having After 1 week, no scales left. 2 applications/ clear margin. day 1 week D34 2 m psoriasis Many red plaques with Back psoriasis: thick scales started to be N/A Age: 30's B ring gel vulgaris attached thick scales having remitted in 3 min after application, Sex: male formulation clear margin presented on almost disappeared after 30 min. 1 application back and neck. Neck psoriasis: erythema and scales remitted in 3 min after application.

TABLE-US-00007 TABLE 7 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D23 2 m cyshidrotic Fingers presented with many Erythema and bullae remitted in 2 min N/A Intractable Age: 30's B ring gel eczema intractable bullae, being after application. After 1 day, erythema with Sex: male formulation dyshidrosis eczematous with erythema and scales remitted and bullae steroid 2 applications/ and cracks. disappeared. ointment. day 1 day D24 2 m miliaria Red small papules and flare Pruritus, erythema and red small N/A Age: 50's B ring gel hyper- with itches. papules remitted after 3 min. Sex: male formulation hidrosis 1 application D25 1 m chronic Many wheals with drastic Pruritus removed just after application. 10 .fwdarw. 0 Age: 50's B ring gel urticaria itches. Wheals started to be gradually remitted Sex: male formulation just after application and disappeared 1 application after 7 min. D26 1 m rosacea Suffering from rosacea for 2 Tautness and tingling itches removed N/A Intractable Age: 30's B ring gel years. 2 months of facial just after application. Diffuse flare, after 2 yrs of Sex: formulation application of a steroid follicular papule and pustule remitted conventional female 1 application ointment exacerbated the after 1 min. After 1 week of 2 therapy symptoms, developing application/day, anti-inflammatory effect without telangiectasia, diffuse flare, was observed. Diffuse flare, follicular satisfactory follicular papule and pustule papule and pustule remarkably therapeutic on cheeks. Subjective improved. Itches and hot flush removed. effect. symptoms include tingling itches and tautness. D35 4 m perioral 2 years of facial application of Diffuse flare and follicular papule N/A Age: 60's B ring gel dermititis a steroid ointment resulted in remarkably remitted 2 min after Sex: formulation circumoral telangiectasia, application. After 2 application/day for 2 female 1 application diffuse flare and follicular days flare improved and scales papule. Subjective symptoms removed. include heat sensation.

TABLE-US-00008 TABLE 8 Pruritus before and after Pre-application symptoms Post-application symptoms application Case Treatment Disease (severity) (severity) (VAS) Notes D36 2 m rosacea Presented with circumoral Diffuse flare and follicular papule Age: 50's B ring gel telangiectasia, diffuse flare remarkably remitted 2 min after Sex: male formulation and follicular papule. application 1 application D37 1 m acne Oily skin accompanied by Seborrhea remitted 2 min after N/A Faster and Age: 20's B ring gel vulgaris suppurative inflammation with application. Anti-inflammatory effect stronger Sex: female formulation many pustules and comedos remitted suppurative inflammation and anti- 1 application/ on lower jaw. pustules on lower jaw. Pustules reduced inflammatory day and dried 3 days after application, and effect than 3 days domedos disappeared. applying/ administrating conventional antibiotics. D38 Right cheek: acne Multiple follicular papules and Anti-inflammatory effect observed 3 min N/A Age: 40's 1 m vulgaris pustules on both cheek. Oily after application, pustules and papules Sex: female B ring gel skin. started to be reduced and flattened, and formulation flare started to be removed. After 1 1 application/ week pustules and papules substantially day remitted and flare further remitted. After 2 weeks 2 weeks pustules, papules and flare almost disappeared. Left cheek: No change in symptoms 3 min after BNP gel application. After switching to B ring single formulation, similar effects as right application, cheek were obtained. then 1 m B ring gel formulation 1 application/ day 2 weeks D39 Right cheek: acne Oily skin accompanied by Anti-inflammatory effect by B ring Age: 10's 2 m vulgaris sppurative inflammation with application was observed on right cheek Sex: male B ring gel multiple red papules and in 3 min. Suppurative inflammation formulation pustules on both cheeks. remitted. 1 application Left cheek: No anti-inflammatory effect on gel formulation gel-applied left side. 1 application D40 4 m acne Seborrheic skin with multiple Comedos faded in 5 min after Age: 10's B ring gel vulgaris comedos and follicular application. Oily skin improved to better Sex: female formulation papules/pustules on forehead. texture. Red papules and pustules 1 application reduced.

[0227] As shown in Tables 1 to 8, in all subjects, or in all dermatitis of any symptoms, emission or elimination of various symptoms of dermatitis was observed at the site where B ring gel formulation was applied.

[0228] Specifically, when B ring gel formulation was applied, remission or elimination of pruritus was observed immediately after the application. Generally in dermatitis, patients tend to scratch where pruritus was felt, potentially causing severer dermatitis. Such a remarkable remission or elimination of pruritus in this manner can consequently prevent aggravation of dermatitis.

[0229] In all cases, the effects of B ring gel formulation were superior to those with BNP gel formulation. Specifically, B ring gel formulation had faster-acting and longer-lasting effects as well as more potent antipruritic effect as compared with BNP gel formulation. It was confirmed that these effect cannot be obtained by the gel base.

3.2 Comparing B Ring Gel Formulation to Gel Base by Two-Sided Application

[0230] For subjects suffering various dermatitis, either B ring gel formulation or the gel base was applied to the affected site, and changes in symptoms before and after the application were observed (Table below).

TABLE-US-00009 TABLE 9 Case Applied site Treatment Disease Pre-application symptoms Post-application symptoms E1 Right B ring atopic Face presented with infiltrating Flare and pigmentation removed 2.5 min after Age: 20's dermatitis erythema, scales and application. Skin elasticity and desiccation Sex: female pigmentation, with intractable improved. Itches eliminated. Left gel base itches No remission of pigmentation, infiltrating erythema, scales and desiccation, with itches left. E2 Right B ring chronic Stiffness and hard lichenification Flare remitted soon after application. After Age: 30's eczema on both forearms, with infiltrating 3 min skin softened and moisturized. Itches Sex: male erythema and desiccation. eliminated. Left gel base No improvement in lichenification, itching and flare after 3 min. E3 Right B ring atopic Presented with hot flush and Flare started to fade soon after application, Age: 10's dermatitis puffiness. and was remarkably remitted after 3 min. Sex: female Left gel base No improvement in flushing after 3 min. E4 Right B ring chronic Stiffness and hard lichenification Flare remitted soon after application, Age: 40's eczema on both forearms, with infiltrating remarkably remitted after 3 min with improved Sex: male erythema, scales and hot infiltration and hot sensation, and softened sensation. skin. Left gel base Infiltration felt 3 min after application. Skin was hard with unimproved flare and hot sensation.

4. Confirming Cosmetic Effect

[0231] Effects of a cosmetic comprising B ring were evaluated from the results obtained using a gel formulations, a bath agent and a hair cosmetic product (shampoo or rinse) or body soap comprising the B ring to confirm their effects.

4.1.1 Confirming Skin-Improving Effect (Comparing B Ring Gel Formulation to BNP Gel Formulation)

[0232] The B ring gel formulation or BNP gel formulation was applied to subjects. The B ring gel formulation was applied on the right cheek or eye of each subject, the BNP gel formulation on the left cheek or eye, and changes in skin conditions were observed and compared (Table below). The changes in skin conditions were determined based on physical sensory evaluation by the subject and our objective observation as physicians.

TABLE-US-00010 TABLE 10 Case Applied site: method Pre-application symptoms Post-application symptoms F1 Right: B ring Presented with wrinkles, cheek 2 min after application, wrinkles on eye corner and forehead were Age: 69 gel formulation flabbiness, pigmentation on eye shallowed, skin was full. After 3 min, cheek flabbiness and skin Sex: female 0.5 mol, corner. texture were improved, and pigmentation on eye corner was less 1 application prominent. After application for 3 days, large wrinkle on cheek was shallowed, flabbiness was improved, skin was firm, resilient, full and moisturized. Left: applied No improvement in wrinkles after 3 min. gel base F2 Right: B ring Presented with wrinkles, cheek 2 min after application, wrinkles at eye corner due to desiccation Age: 45 gel formulation flabbiness and desiccation. was less prominent, skin texture was improved, moisturized and Sex: female 0.5 mol, softened. 1 application Left: applied No improvement in wrinkles and desiccation after 2 min. gel base F3 Right: B ring Presented with desiccation, light 30 sec after application, skin desiccation was improved, skin was Age: 21 gel formulation erythema, tautness and itch. moisturized and flexible with no tautness. After 1 min, flare was Sex: female 2 mol, remitted and itches were removed. After 4 min, further softness was 1 application given and desiccation was remarkably improved. Left: applied No improvement observed in desiccation, itching and flare after gel base 4 min. F4 Right; B ring Presented with desiccation and 3 min after application, skin desiccation was improved, skin was Age: 24 gel formulation light erythema. moisturized and firm, with improved texture. Skin was flexible, Sex: female 2 mol, soft and moisturized. 1 application Left: applied No improvement observed in desiccation and flare after 3 min. gel base F5 Right: B ring Presented with wrinkles on both 2 min after application, wrinkles on both corners of eyes were less Age: 28 gel formulation corners of eye, and desiccation. prominent, cheek dryness was improved, skin was moisturized and Sex: female 2 mol, flexible. Perioral desiccation was remitted and skin became full 1 application and flexible. Left: applied No improvement observed in desiccation and wrinkles after 2 min gel base at gel-applied site on the left. F6 Right: B ring Presented with skin desiccation, 1 min after application, itches were remarkably remitted. After Age: 30 gel formulation oily skin and red papules around 3 min, skin desiccation and flare were improved, and skin was Sex: female 2 mol, nose, and light erythema and moisturized and flexible with improved texture. Red papules 1 application rough skin texture. around nose almost disappeared, combination state of desiccation and oily skin was improved, and skin is kept healthy. Left: applied 3 min after application, desiccation, flare and itches are left, gel base and seborrhea and papules around nose were not remitted.

[0233] Table below summarizes examples in which B ring gel formulation was applied without applying two-sided. The changes in skin conditions were determined by similar method as described above.

TABLE-US-00011 TABLE 11 Case Method of application Pre-application symptoms Post-application symptoms G1 B ring gel formulation Presented with cheek flare, open After B ring application, flare was repressed, skin Age: 50 1 mol pores, rough and hard skin texture. texture was improved, skin was moisturized and softened. Sex: female 1 application G2 B ring gel formulation Presented with wrinkles and 3 min after application at B ring-applied site, wrinkles Age: 49 2 mol desiccation on forehead and eye on forehead and eye corner were shallowed and less prominent, Sex: female 1 application corner. skin desiccation was improved, and skin was moisturized and flexible. G3 B ring get formulation Presented with male skin 1 day after application at ring-applied site, skin desiccation Age: 40's 1 mol desiccation and roughness. and roughness were improved, and razor burn was prevented. Sex: male 1 application G4 B ring gel formulation Presented with miliaria and 3 min after application at B ring-applied site, miliaria was Age: 50's 2 mol pruritus accompanied with flare. prevented, flare and itches were remitted. Sex: male 1 application

[0234] Accordingly, it was shown that B ring of the present invention has a skin moisturizing effect and is capable of improving skin texture, and that it is also effective in improving skin, for instance in lightening (spots, dullness) and anti-aging (flabbiness, resilience, large wrinkles) as well as in improving mucosal condition such as roughened lips.

[0235] In all subjects, a remarkable improving effect in skin and its persistence was observed at the site where B ring gel formulation had been applied as compared to where BNP gel formulation had been applied. Specifically, effects as follows were observed at the site where B ring gel formulation had been applied as compared to where BNP gel formulation had been applied with significance: effects of improving skin, moisturizing skin, improving skin texture, providing skin with resilience and firmness, softening skin, suppressing skin desiccation, fading fine wrinkles, supplying and keeping skin with water and oil, shallowing and diminishing crow's feet, eliminating skin rawness, eliminating and suppressing itches, relieving skin roughness and maintaining skin condition healthy.

[0236] Particularly, in subjects in their 20's or 30's, remarkable effects such as improving or eliminating wrinkles, improving cheek flabbiness, and supplying and keeping skin with moisture, water and oil, and the effects such as giving skin firmness and tightness, lifting up cheeks, eliminating skin rawness and relieving skin roughness were observed at the site where B ring gel formulation had been applied as compared to the site where BNP gel formulation had been applied, and these effects were greater as compared to those at BNP gel-applied site.

[0237] Moreover, in subjects in their 40's and 50's, remarkable effects such as improving flabbiness, large wrinkles and desiccation, giving skin firmness, fading dullness and spots were exhibited immediately after the application of B ring gel formulation, and these effects were greater as compared to those upon applying BNP gel formulation.

[0238] The effects exhibited significantly faster in B ring gel formulation as compared to BNP gel formulation. Specifically, effects were confirmed in most cases such that wrinkles were vanished or started to vanish and faded and skin was made full and resilient. These effects lasted for at least several hours thereafter. No irritating symptoms were observed with application of B ring gel formulation.

[0239] FIG. 1 shows a photograph of the right-side face of a subject in her 60's after the application of B ring gel formulation. In this subject, effects were observed at 20 minutes after the application of B ring gel formulation such that the skin were moist, provided with moisture, complexion and softness, given firmness, improved texture, provided with resilience, and large wrinkles faded. This result was compared to BNP gel formulation as a comparative example, indicating that the cyclic peptide of the invention is not only effective on fine wrinkles but also is effective on wrinkles and flabbiness, and that it is also effective in fading spots and dullness.

4.1.2 Confirming Skin-Improving Effect (Comparing B Ring Gel Formulation to Gel Base)

[0240] In order to examine the presence or absence of the contribution by the gel base to the effects of B ring gel formulation and BNP gel formulation, three subjects were administered B ring gel formulation on their right face and the gel base on the left, and changes in skin conditions on the subject's faces were observed (Table below).

TABLE-US-00012 TABLE 12 Case Applied site: method Pre-application symptoms Post-application symptoms H1 Right: B ring gel Rough, stiff and dry skin texture. After 2 min skin was left soft. Age: 48 formulation Sex: female Left: applied gel base Skin was tingling. Open pores and rough skin texture were not improved. H2 Right: B ring Sensitive dry skin with roughness 1 min after application, skin tingling sensation Age: 26 and tingling stopped. After 2 min, B ring-applied side was felt Sex: female more moisturized. Left: applied gel base 1 min after application, skin was still tingling. After 2 min, the subject left that moisturizing effect is weak. H3 Right: Sensitive skin with tingling. Age: 30 Left: applied gel base Skin was tingling and temporarily red. Skin desiccation Sex: female was not improved. White scales were observed.

[0241] As a result, all subjects felt tingling sensation at the site where the gel base was applied, and dry feeling of skin was not improved. Also, in all subjects, no effect (e.g., improved skin texture or closed pores) was observed at the site where the gel base was applied. On the other hand, in all subjects, the skin area on which B ring gel formulation was applied has been moisturized and softened.

4.2 Observation on Mucosal Condition Upon Application of B Ring Gel Formulation.

[0242] The B ring gel formulation was applied to normal subjects who had symptoms of roughened lips, and changes in mucosal condition were observed (Table below). The changes in mucosal conditions were determined based on physical sensory evaluation by the subject and our objective observation as physicians.

TABLE-US-00013 TABLE 13 Case Applied site: method Pre-application symptoms Pos-application symptoms I1 B ring 1 mol Cracks on mouth corner with pain. Skin became moisturized soon after application. After 2 min, Age: 20's 1 application Desiccation and wrinkles of lips. cracks on mouth corner were remitted. After 5 min, lips were Sex: female soft and no longer felt dry or tingling. I2 B ring 0.5 mol Annoying lip desiccation. 3 min after application, lips became moisturized. Age: 30's 1 application Sex: female I4 B ring 2 mol Annoying lip desiccation. After 2 min, lips were full and soft, and wrinkles were less Age: 30's 1 application prominent. Moisturized. Sex: female I3 B ring 2 mol Lips were dry and tingling, After 2 min, lips were full, soft and moisturized, and wrinkles Age: 30's 1 application with wrinkles and scales were less prominent. Tingling sensation stopped. Sex: female I4 Right: B ring Cracks on mouth corner with pain. 2 min after application, cracks on mouth corner were remitted Age: 20's 1 mol Dry and roughened lips with no more pain, and moisturized. Sex: male 1 application Left: gel base 2 min after application, cracks on mouth corner and pain were 1 application not improved

[0243] Accordingly, it was confirmed that the cyclic peptide of the invention has an activity of improving mucosal condition and it also has an immediate effect in such activity.

4.3 Observation on Scalp/Hair Condition Upon Application of B Ring Gel Formulation.

[0244] The B ring gel formulation was applied to normal subjects who had symptoms of scalp/hair, and changes in conditions of scalp and hair were observed (Table below). The changes in scalp/hair were determined based on physical sensory evaluation by the subject and our objective observation as physicians.

TABLE-US-00014 TABLE 14 Applied site: Case method Pre-application symptoms Post-application symptoms J1 B ring 0.5 mol Presented with desiccation and After 2 min. itches were removed, scalp was relieved from Age: 40's 1 application itches of head. roughness and desiccation and moisturized. After that scalp Sex: male was prevented from desiccation and kept moisturized. J2 B ring 1 mol Presented with erythema and itch Hair growth was stimulated, and an increase in hair volume Age: 50's 1 application/day, of head, and thinning and falling of was observed, and thinning of hair was improved. Hair was Sex: female 3 weeks hair. provided with moisture, flexibility and radiance. J3 B ring 1 mol Presented with thinning and falling A decrease in falling hair was observed, showing an effect Age: 40's 1 application/day, of hair. of preventing alopecia. Hair growth was stimulated, and an Sex: female 5 days increase in hair volume was observed, and thinning of hair started to be improved. Hair was provided with moisture, flexibility and radiance. J4 B ring 2 mol Presented with seborrheic dandruff After 2 min, itches were removed, scalp seborrhea and Age: 40's 1 application and itches. dandruff started to be improved. After that scalp became Sex: female and was kept clean. J5 B ring 2 mol Presented with desiccation, mild After 1 min. itches were removed. After 2 min, scalp Age: 20's 1 application erythema and itches. seborrhea and dandruff started to be improved. After 4 min, Sex: male erythema and skin roughness of scalp was improved. After that skin roughness was prevented. J6 B ring 2 mol Presented with scalp seborrhea, After 2 min, itches were removed, scalp seborrhea was improved. Age: 60's 1 application dandruff and itches. After that dandruff was removed, scalp became healthy and clean. Sex: male J7 B ring 1 mol From the beginning of 5th month of Upon being applied to alopecia 5 month post partum, an effect Age: 30's To decalvant site 3rd pregnancy, significant hair loss was exhibited on hair loss in eyebrows and head where hair Sex: female on head and brows in eyebrows, lashes and scalp growth was stimulated, improving thinning of hair. of subject: 1 occurred, loosing almost all hair. application/day post partum, continued

[0245] Accordingly, BNP cyclic peptide of the present invention can provide and keep scalp and hair with moisture, give them moderate water and oil, improve and prevent desiccation, and clean scalp and hair. It is further effective for suppressing itch and dandruff of scalp and also effective for preventing thinning or loosing hair, for growing hair and promoting or stimulating hair growth, and forcing hair growth, and furthermore effective for an improvement in alopecia after illness or postnatal alopecia.

4.4 Observation on Skin Condition Upon Using Bath Agent Comprising B Ring

[0246] Subjects were given bath in 37 to 41 C. hot water comprising B ring dissolved at 0.01 M (100 ml of 20 M B ring solution dissolved in 200 L hot water) once a day for 14 days, and skin condition of the subjects was compared to the case of hot water alone (Table 14). The changes in skin conditions were determined based on physical sensory evaluation by the subject and our objective observation as physicians.

TABLE-US-00015 TABLE 15 Pre-application symptoms ( , cracks, chaps Case Treatment and eczema) Post-application symptoms Changes in subjective symptoms K1 B ring Eczema Gradual remission of itches after bath was remitted. Age: 30's addition erythema and skin Skin was moisturized. Sex: female desiccation. K2 Hot water Eczema Flare and desiccation Enhanced itches and desiccation Age: 30's exacerbated. after bath. Sex: female K3 BNP Eczema No remission of skin No subjective remssion of skin Age: 30's addition desiccation or erythema. desiccation at this concentration. Sex: female K4 B ring Chaps, rough skin Remission of desiccation. Remission of itch and pain. Skin Age: 60's addition of hands, itches became smooth Sex: female with tingling K5 Hot water Chaps and rough Exacerbation of desiccation. Exacerbation of itches and Age: 60's skin of hands tingling after some time after Sex: female bath. K6 B ring and itches Remission of Remission of itches and improvement Age: 20's addition in skin texture. Sex: male K7 Hot water and itches No improvement in No remission of itches. Age: 20's Sex: male K8 B ring Cracks and pain Cracks gradually became Relief of pain. Skin was moisturized. Age: 40's addition on sore shallower and improved Sex: female upon application K9 Hot water Cracks and pain No improvement in cracks. No remission of pain. Age: 40's on sore Sex: female indicates data missing or illegible when filed

[0247] Accordingly, it was confirmed that the bath agent comprising B ring of the present invention has an ameliorating effect on miliaria, cracked or chapped skin, and also an improving effect on eczema. Furthermore, it was shown to improve desiccation, pain and itch of the skin, indicating that it is effective for improvement of skin condition.

4.5 Observation on Conditions of Scalp and Hair or Skin Upon Using Hair Cosmetics (Shampoo, Rinse) or Body Soap Comprising B Ring

4.5.1 A Shampoo Comprising B Ring was Mixed and Prepared in Composition as Follows.

[0248]

TABLE-US-00016 TABLE 16 Name of ingredient (tradename) Content (%) A Sodium cocoylmethyltaurine solution 10.0 (NIKKOL CMT-30) Sodium laureth lactate (NIKKOL SBL-2N-27) 20.0 Lauryl betaine solution (NIKKOL AM-301) 10.0 Cocamide DEA 4.0 Antiseptic q.l. B Citric acid 0.1 Propylene glycol 2.0 Guar hydroxypropyl trimonium chloride 0.5 Water to fill up 100.0 C Water, B ring (B ring concentration 1 M) 1 mL

[0249] A and B were dissolved with heat at 70 C. To A, B was added, stirred and mixed. C was added at 40 to 35 C. while being further stirred, and allowed to cool to room temperature as kept being stirred.

4.5.2 A Conditioner Comprising B Ring was Mixed and Prepared in Composition as Follows.

[0250]

TABLE-US-00017 TABLE 17 Name of ingredient (tradename) Content (%) A Pentylene glycol 1.50 BG 3.50 Glycerin 1.00 Methyl paraben 0.20 Carbomer 0.20 EDTA-2Na 0.10 Water to fill up 100.0 B Composite emulsifier (NIKKOL Nikkomulese LC) 4.00 Methyl heptyl laurate 3.50 Squalane 0.50 Cetearyl alcohol 1.50 Macadamia nut oil 0.50 Avocado oil 0.50 Shea oil 0.50 Propyl paraben 0.10 C Water, B ring (B ring concentration 1 M) 1 mL

[0251] A and B were dissolved with heat at 80 C. Then A was stirred by a homomixer while B was gradually added thereto, and the mixture was emulsified. C was further added to the mixture, and the mixture was allowed to cool to 35 C. as kept being stirred.

4.5.3 A Body Soap Comprising B Ring was Mixed and Prepared in Composition as Follows.

[0252]

TABLE-US-00018 TABLE 18 Name of ingredient (tradename) Content (%) A Cocoyl glutamic acid TEA solution 30.0 Sodium trideceth-4 carboxylate 5.0 (KIKKOL ECTD-3 NEX) Sodium cocoamphoacetate solution 10.0 (KIKKOL AM-101) PEG-50 hydrogenated castor oil (KIKKOL HCO-50) 0.5 1,3-butylene glycol 5.0 Antiseptic q.l. B EDTA-2Na q.l. Water to fill up 100.0 C Water, B ring (B ring concentration 1 M) 1 mL

[0253] A and B were dissolved with heat at 80 C. Then B was added to A while being stirred. C was further added to the mixture at 40 to 35 C. while kept being stirred, and the mixture was allowed to cool to room temperature as kept being stirred.

[0254] Subjects used shampoo, rinse or body soap prepared as above once a day for 14 days, and resulted conditions of scalp and hair or skin of the subjects were assessed (Table below). Changes in skin condition was assessed by dandruff of scalp. Changes in erythema was assessed by visual observation by the subject. Itches, moisturized feeling, combing, glow, resilience and elasticity of hair were assessed by subject's physical sensory evaluation.

TABLE-US-00019 TABLE 19 Case Treatment Pre-application symptoms Post-application symptoms Subjective changes L1 shampoo/ Presented with erythema and Remission of scalp Improvement in itches. Hair was Age: 60's treatment itches on scalp, and thinning erythema and a decrease moisturized and provided with Sex: male and falling hair. in hair falling. resilience, radiance and elasticity. L2 shampoo Presented with desiccation Improvement in dandruff Improvement in itches. Hair was Age: 20's and dandruff on scalp and desiccation of scalp. moisturized and provided with Sex: female accompanied with itches resilience, radiance and elasticity. Improved. With better combing. L3 body soap Dry and sensitive skin. Improvement in dry skin, improvement in itches after bath. Age: 60's relief of itching after bath. Sex: female L4 body soap Dry and sensitive skin. Improvement in dry skin, Improvement in itches after bath. Age: 60's relief of itching after bath. Sex: female L5 body soap Dry skin. Improvement in dry skin. Skin was moisturized after bath. Age: 20's Sex: male

[0255] Accordingly, the use of the hair agent comprising B ring of the present invention had an fast-acting improving effect on itch, erythema and desiccation of scalp. Moreover, it was shown to be effective in improving symptoms of thinning and falling hair when it was used continuously for at least 2 weeks. Furthermore, it was shown that the use of the body soap comprising B ring of the present invention has an improving effect on dry or sensitive skin.

5. Confirming Therapeutic Effect on Alopecia

[0256] Either B ring gel formulation or BNP gel formulation was applied, and their effects on symptoms associated to various alopecia were observed (Table below). The changes in symptoms were determined based on physical sensory evaluation by the subject and our objective observation as physicians.

TABLE-US-00020 TABLE 20 Case Treatment Disease Post-application symptoms A21 Right: B ring 1 mol Male alopecia 7 weeks after application, hair became elastic and thick, Sex: male 1 application/ a drastic decrease in hair-falling Consequently, hair became Age: 50's day, continued. dense and thinning less prominent B ring-containing gel Left: BNP gel formulation exhibited more remarkable hair growing affect formulation 1 application/ day, continued. A5 B ring gel Male alopecia and 2 weeks after starting applying B gel formulation, the Sex: male formulation alopecia existing soft hair in parietal area became thicker and Age: 50's 1 application/ accompanied with longer and turned black.. Hair got resilient and alestic. day onto head dandruff. Previous Newly grown terminal hairs remarkably decreased thinning decalvent site, use of -containing area. Dandruff was suppressed overall, particularly in continued. drug caused strong parietal area. was eliminated. irritation, itches and erythema to the subject and its use was terminated A22 B ring gel Seborrhoic alopecia 2 weeks after application, thinning or hair was improved. Sex: female formulation accompanied with exhibiting no more showing-through of scalp confirming Age: 40's 2 application/ dandruff. hair-growing stimulation. Dandruff stopped, scalp seborrhea day onto was improved and pruritus was removed. Furthermore, scalp parietal and skin became clean. decalvant site. indicates data missing or illegible when filed

TABLE-US-00021 TABLE 21 A10 B ring gel formulation Alopecia universalis. 2 weeks after starting application, on both sides of head, Sex: male 0.5 mol Alopecia on whole head. regeneration of pores and growth of terminal hair were Age: 20's BNP gel formulation Previous therapy with confirmed, which were more remarkable and hair growth area 0.5 mol carpronium chloride was wider on the B ring gel formulation-applied right side. and steroid pulse first Hair growth area kept increasing to the whole head. The exerted hair growth- density, elongation rate and thickness of hair were greater stimulating effect, but on the B ring gel formulation-applied right side. Upon caused complete hair increasing the concentration of application to 1 mol, loss after 1 month. rate of growth and elongation were increased. A11 Right side of head decalvant Ophiasis and multiple Right forehead ophiasis site: 7 days after starting applying Sex: female site: B ring gel formulation alopecia. B ring gel formulation, a hair growth stimulation effect was Age: 40's 1 application/day for 1 week. observed and hair became thicker terminal hair, hair-growing 1 or 2 applications/week for 1 area kept increasing. year thereafter. Left BNP gel formulation-applied site: few soft hair observed, Left side of head decalvant whose growth was slow and in obviously smaller area. site: BNP gel formulation After continuing 1 or 2 applications/week for 1 year: 1 application/day for 1 week. Right forehead B ring gel formulation-applied site: hair- 1 or 2 applications/week for 1 growing area kept increasing and hair elongation rate was year thereafter. fast (FIG. 2). Left BNP gel formulation-applied site: hair growth was observed though its area was smaller as compared to the right side, and majority of hairs was thin and soft (FIG. 2).

TABLE-US-00022 TABLE 22 A12 Head decalvant site: Alopecia areata 1 week after application, remarkable stimulation and growth Sex: female B ring gel formulation of terminal hair was observed. Age: 20's 1 application/day, continued. A12 Subject's head decalvant Alopecia areata 1 week after application, remarkable stimulation, growth Sex: female site: B ring gel formulation and elongation of terminal hair was observed. Age: 30's 1 application/day, continued. A24 Right half of head decalvant Drug-induced Pre-application: 1 month after termination of anticancer Sex: female site: B ring gel formulation alopecia agent therapy. Bilateral severe diffuse alopecia on whole Age: 50's (1 mol), 2 applications/ head. Thinning of hair on whole head. Hair was short and day for 2 weeks thin, mostly white. Left half of head decalvant 1 day after application at B ring-applied site, subjective site: BNP gel formulation hair elongation and growth stimulation were observed, and (1 mol), 2 applications/ grown hairs were predominantly black (non-white). After 2 day for 2 weeks weeks, hair growth was observed at both sides. Hair were elongated with less falling hairs. Hair elongation rate is faster on B ring-applied site. Hairs are also thicker and denser, and contain more non-white hairs as compared to BNP-applied site. 3 weeks after application, remarkable growth of terminal hairs was observed.

5.2 Case Summary

[0257] In the cases of the subjects having female pattern alopecia, male pattern alopecia, alopecia universalis, ophiasis or alopecia areata multilocularis as described above, a significant decrease in falling hair was observed, and the area of terminal hair growth was expanded, and the growth of the terminal hair was also faster, when B ring gel formulation of the working example was applied compared with BNP gel formulation as a comparative control (See, FIG. 2). Addingly, the existing hairs obtained resilience and their elasticity was increased. In all cases, symptoms were significantly improved as compared to the cases when BNP gel formulation of the comparative example was applied. Accordingly, B ring gel formulation had superior effects on the symptoms described above as compared to BNP gel formulation.

[0258] In addition, the B ring gel formulation could improve seborrhea, clean scalp and suppress dandruff. It also prevents hairs from turning white, grows black hairs, and improves thinning of hair. Moreover, it exhibits an antipruritic effect within 10 minutes, demonstrating an excellent immediate effect. It could suppress ithes within 3 minutes after application in some cases.

6. Confirming Therapeutic Effect on Rhinitis

6.1 Cases

[0259] Either ring nasal drop, BNP gel nasal drop was sprayed, and their effects on rhinitis in the subject was observed (Table 17). The changes in symptoms were determined based on physical sensory evaluation by the subject and our objective observation as physicians.

TABLE-US-00023 TABLE 23 Case Treatment Disease Post-application symptoms (severity) R1 Right nasal cavity: Perennial chronic Right nasal cavity (B ring nasal drop-treated side): Subject felt breath goes through right Sex: female B ring nasal rhinitis and nasal cavity soon after application. After 30 sec, air goes through nasal cavity. After 2 Age: 20's formulation 0.1 ml, rhinorrhea. 20 or min. nose was clear and the subject could breathe comfortably. After 3 min, felt fresh 1 spray more blow/day and retained nasal discharge was removed. No discharge by blowing. A day after Left nasal cavity: application, no discharge from right nasal cavity. BNP nasal Left nasal cavity (BNP nasal drop-treated side): 3 min after application, the discharge formulation 0.1 ml, still retained in nasal cavity and drained by blowing. A day after application, mild nasal 1 spray obstruction and rhinorrhea symptoms in left nasal cavity with small amount of retained discharge. R2 Right nasal cavity: Rhinitis with severe Right nasal cavity (B ring nasal drop-treated side): Subject felt breath goes through right Sex: female B ring nasal nasal obstruction nasal cavity soon after application. After 30 sec, obstruction was improved, subject's Age: 30's formulation 0.1 ml, and feeling heavy. suffering in breathing was reduced in right nasal cavity. After 1.5 min, suffering in 1 spray Right nasal cavity breathing was removed in right nasal cavity and breath went though. A day after Left nasal cavity: has severer application, still no obstruction or rhinorrhea. Ths subject felt no irritation upon treatment BNP nasal obstruction symptom. in right nasal cavity formulation 0.1 ml, The subject feels dry Left nasal cavity (BNP nasal drop-treated side): 30 sec after application, obstruction of 1 spray and tingling by left nasal cavity was slightly improved, though the subject did not feel breath goes steroid nasal drops. through as in the right cavity. A day after application, obstruction or rhinorrhea in left nasal cavity were removed. The subject felt mild dryness in left nasal cavity. R3 Right nasal cavity: Perennial allergic Right nasal cavity: Subject felt breath goes through nasal cavity 1 min after application. Sex: female B ring nasal rhinitis with After 3 min nasal obstruction was improved, the subject could breathe through nose. Age: 20's formulation 0.1 ml, rhinorrhea and nasal After 5 min the subject could breathe comfortably through nose, with no nasal discharge 1 spray obstruction. by blowing. Right nasal cavity has severer obstruction with retention of nasal discharge.

TABLE-US-00024 TABLE 24 R4 Right nasal cavity: Chronic rhinitis with Right nasal cavity (B ring nasal drop-treated side): 1 min after treatment, nasal Sex: female B ring nasal severe rhinorrhea and obstruction was improved to some extent. After 2 min, obstruction was improved, the Age: 30's formulation 0.1 ml, nasal obstruction. subject could breathe through nose as usual, and rhinorrhea stopped. After 3 min, no 1 spray Application of steroid nasal discharge from the right nasal cavity by blowing. Left nasal cavity: nasal drops could not Left nasal cavity (BNP nasal drop-treated side): 1 min after treatment, nasal discharge BNP nasal improve symptoms when was retained in the left nasal cavity with no improvement in obstruction After 2 min, formulation 0.1 ml, they were severe. obstruction was improved to some extent, though discharge was retained, showing 1 spray rhinorrhea symptom. After 3 min, nasal discharge was not drained by blowing from left nasal cavity due to obstruction. After 8 min, nasal discharge was drained by blowing. R5 Right nasal cavity: Allergic rhinitis with Right nasal cavity (B ring nasal drop-treated side): 30 sec after treatment, obstruction Sex: female B ring nasal rhinorrhea as main was improved and the subject could breathe through nose. After 2 min, the subject Age: 20's formulation 0.1 ml, symptom and mild could easily breathe through nose. After 8 min, no nasal discharge was drained by 1 spray obstruction. blowing from the right nasal cavity. A day after treatment (after 26 hours), nasal Left nasal cavity: discharge was controlled, and breathing was easier through the right nasal cavity as BNP nasal compared to the left. formulation 0.1 ml, Left nasal cavity (BNP nasal drop-treated side): 2 min after treatment, obstruction was 1 spray still observed. After 8 min, small amount of discharge was drained by blowing. A day after treatment (after 26 hours), nasal discharge was controlled. R6 Right nasal cavity: Allergic rhinitis Right nasal cavity (B ring nasal drop-treated side): nasal obstruction was slightly Sex: female B ring nasal with both rhinorrhea improved soon after treatment. After 1 min, the subject could easily breathe though Age: 20's formulation 0.1 ml, and nasal obstruction nose. After 4 min, no nasal discharge was drained by blowing from the right nasal cavity. 1 spray having constant Nasal obstruction was resolved. After 10 min, pruritus was removed. A day after Left nasal cavity: obstruction and treatment (after 28 hours), no obstruction or rhinorrhea was observed. BNP nasal severe rhinorrhea. Left nasal cavity (BNP nasal drop-treated side): 1 min after treatment, nasal obstruction formulation 0.1 ml, 10-20 blow/day. was not improved. After 3 min, obstruction was still not improved and rhinorrhea was 1 spray Regular dose of oral observed. After 3 min, small amount of discharge was drained by blowing. After 10 min, anti-allergic drug obstruction was slightly improved but not resolved. After 2 hours, obstruction was did not sufficiently resolved and the subject could breathe though nose. A day after treatment (after 28 improve nasal hours), no obstruction or rhinorrhea was observed. obstruction and rhinorrhea. Steroid nasal spray causes mucosa pain.

TABLE-US-00025 TABLE 25 R21 Right nasal cavity: Rhinitis with continuous rhinorrhea Right nasal cavity (B ring nasal drop-treated side): Sex: female B ring nasal and sneeze. Nasal discharge was 30 sec after treatment, the subject could easily breathe Age: 20's formulation 0.1 ml, drained by facing down. through nose. After 1 min rhinorrhea stopped. No nasal 1 spray discharge by facing down. R8 Right nasal cavity: Chronic allergic rhinitis (with both Right nasal cavity (B ring nasal drop-treated side): Sex: female B ring nasal rhinorrhea and obstruction). The 1 min after treatment, obstruction was improved and the Age: 20's formulation 0.1 ml, subject regularly feels nasal subject could easily breathe through nose. The subject 1 spray + 1 spray obstruction. Application of steroid felt no irritation by treatment. The effect lasts for 1 20 min later nasal formulation causes the week thereafter, and rhinorrhea arid obstruction were Left nasal cavity: subject strong irritation. controlled. No nasal discharge by blowing. BNP nasal Accompanied with rhinorrhea and Commercial steroid nasal formulation caused a sharp formulation 0.1 ml, eye pruritus. irritation and recurrence of symptoms within 2 hours. 1 spray + 1 spray Left nasal cavity (BNP nasal drop-treated side): 20 min later R9 Right nasal cavity: Allergic rhinitis (with rhinorrhea). Right nasal cavity (B ring nasal drop-treated side): Sex: female B ring nasal The subject has rhinorrhea all day 2 min after treatment, the subject could breathe through Age: 30's formulation (1 mol) and usually blows 20 or more times nose, with no irritation in the right nasal cavity. 0.1 ml, 1 spray a day. Application of steroid nasal After 4 min the subject could breathe through nose, with Left nasal cavity: formulation causes the subject no nasal discharge drained. BNP nasal strong dry feeling on mucosae of Left nasal cavity (BNP nasal drop-treated side): formulation (1 mol) nasal cavity and pharynx. The 2 min after treatment, obstruction was improved and the 0.1 ml, 1 spray effect of the steroid nasal subject could breathe through nose. After 4 min, in the formulation lasts only 3-4 hours, left nasal cavity, the subject drained small amount of and after the effect ended, nasal discharge by blowing. In the right nasal cavity, sneezing and nasal discharge last the effect of the nasal treatment lasted for 4 hours. for several hours and need Blowing caused obstruction in the left nasal cavity but not frequent blowing. in the right. The subject felt that B ring nasal formulation is much better to the steroid nasal formulation regarding the effect on the symptoms, long-lasting and fast-acting effect, lower frequency of use required, and absence of irritating symptom upon use.

TABLE-US-00026 TABLE 26 R10 Right nasal cavity: Allergic rhinitis with rhinorrhea Right nasal cavity (B ring nasal drop-treated side): Sex: male B ring nasal and obstruction complicated with 2 min after nasal treatment, obstruction was improved Age: 30's formulation (1 mol) rhinosinusitis. Obstruction in the and the subject could breathe through nose. The subject 0.1 ml, 1 spray right nasal cavity due to could breathe more comfortably in the right nasal cavity Left nasal cavity: rhinosinusitis is severer than the as compared to the left. The subject felt no irritation BNP nasal left and could not be improved by in the right nasal cavity. After 4 min, the subject could formulation (1 mol) steroid nasal formulation, making very easily breathe, with no nasal discharge drained by 0.1 ml, 1 spray the subject incapable of breathing blowing. through nose. If left untreated, Left nasal cavity (BNP nasal drop-treated side): nasal discharge is drained from 4 min after nasal treatment, nasal obstruction was not nostrils spontaneously. resolved. Nasal discharge was decreased as compared to pre-treatment, though draining was still observed. R11 Right nasal cavity: Perennial rhinitis with rhinorrhea Right nasal cavity (B ring nasal drop-treated side): Sex: male B ring nasal and obstruction complicated with 2 min after nasal treatment, the subject could breathe Age: 20's formulation 0.1 ml, rhinosinusitis, having severe through nose. After 5 min, nasal discharge stopped, 1 spray obstruction. Very high frequency enabling the subject to breathe though nose. The subject Left nasal cavity: of blowing. These symptoms were felt no obstruction and was able to breathe through nose BNP nasal not improved by any previously throughout 1 week after nasal treatment. formulation 0.1 ml, used nasal formulation. Left nasal cavity (BNP nasal drop-treated side): 1 spray 5 min after nasal treatment, obstruction was not resolved, with nasal discharge present in the left nasal cavity. After 10 min. obstruction was not resolved, making the subject incapable of breathing through nose.

6.2 Case Summary

[0260] In all cases described above, both rhinorrhea and nasal obstruction were quickly improved or eliminated when B ring nasal drop of the working example had been applied as compared to the case when BNP nasal drop of comparative example had been applied. Accordingly, B ring nasal drop had faster effect than BNP nasal drop. Addingly, the effect of B ring nasal drop was more than equal to that of BNP nasal drop, and because the effect lasted for longer time period, it can suppressing recurrence of symptoms.

[0261] The following table summarizes information from the cases described above, within the range recognizable at the time when symptoms of nasal obstruction and rhinorrhea were improved.

TABLE-US-00027 TABLE 27 No resolution within within within within 3 min 5 min 10 min 10 min Resolution of rhinorrhea B ring nasal formulation 3 4 1 0 BNP nasal formulation 0 0 0 7 Resolution of nasal obstruction B ring nasal formulation 6 5 0 0 BNP nasal formulation 0 0 0 8

7. Analysis of Binding State of Human BNP and Type A Receptor

[0262] The superior pharmacological effects of the B ring-compound of the invention on various diseases relative to conventional BNP has been explained so far with reference to data based on respective clinical cases. Such pharmacological effects is also supported by the result of the conformation analysis of the compound performed by the inventors, which is explained by following experimental report for reference.

[0263] In order to investigate binding state of human BNP (BNP-32) and its receptor Type A receptor (NPR-A), an in silico analysis was performed by homology modeling using conformation. Swiss-Pdb viewer and SWISS-MODEL were used for modeling.

7.1 Template Structure

[0264] Firstly, prior to the above analysis, a template structure for investigation on binding state of human BNP and Type A receptor was selected. As this template structure, the conformation of the complex of rat NPR-A and rat ANP peptide (PDB ID: 1T34) were used. Rat NPR-A is a homodimer composed of A and B strands. The conformation of such rat NPR-A has been determined by X-ray crystalline structural analysis such that 21 residues from Cys7 to Arg 27 of rat ANP were bound. The conformation of rat NPR-A was obtained from the database of protein conformation, Protein Data Bank. Amino acid homology between human NPR-A and rat NPR-A is 85%.

7.2 BNP Peptide Model

[0265] In the present study, as BNP peptide model, a region from Cys10 to Arg30 in human BNP (BNP-32, SEQ ID NO: 13, Ser-Pro-Lys-Met-Val-Gln-Gly-Ser-Gly-Cys-Phe-Gly-Arg-Lys-Met-Asp-Arg-Ile-Ser-Ser-Ser-Ser-Gly-Le u-Gly-Cys-Lys-Val-Leu-Arg-Arg-His), i.e., an amino acid sequence (SEQ ID NO: 14, Cys-Phe-Gly-Arg-Lys-Met-Asp-Arg-Ile-Ser-Ser-Ser-Ser-Gly-Leu-Gly-Cys-Lys-Val-Leu-Arg) was used. In the peptide model above, the amino acid sequence of B ring of the invention corresponds to a region from Cys10 to Cys26 of human BNP above, i.e., Cys-Phe-Gly-Arg-Lys-Met-Asp-Arg-Ile-Ser-Ser-Ser-Ser-Gly-Leu-Gly-Cys (SEQ ID NO: 15).

7.3 Homology Modeling

[0266] In order to speculate the amino acid residues that are involved in binding of human NPR-A and human BNP, a complex model in which BNP peptide is bound to human NPR-A was constructed by homology modeling. Specifically, human BNP peptide model was constructed based on the template structure of rat ANP peptide, and human NPR-A model structure based on the template structure of rat NPR-A.

[0267] Next, residues that are involved in the interaction was speculated from the amino acid residues detected between human BNP peptide model and human NPR-A.

[0268] The results showed that BNP peptide model is bound to human NPR-A dimer being sandwiched between the A strand and B strand. Amino acids in the amino acid sequence, e.g., the amino acid sequence used as the model (SEQ ID NO: 14, Cys-Phe-Gly-Arg-Lys-Met-Asp-Arg-Ile-Ser-Ser-Ser-Ser-Gly-Leu-Gly-Cys-Lys-Val-Leu-Arg) are expressed as amino acid (number). Namely, the number in parentheses denotes the relative position counted from the N-terminal of the amino acid sequence, for example, Phe2 indicates an amino acid that is the second Phe counted from the N-terminal of the peptide model.

[0269] In the constructed complex model, the presence of a hydrophobic bond by Phe2 side chain, a hydrogen bond by Phe2 main chain, and hydrogen bonds by side chains of Arg4, Met6, Arg8, Ser10 and Ser11 were speculated between BNP peptide model and the A strand of NPR-A. Phe2, Arg4, Met6, Arg8, Ser10 and Ser11 in BNP peptide model correspond to Phe11, Arg13, Met15, Arg17, Ser19 and Ser20 in human BNP respectively. Namely, this result suggests that these amino acid residues in human BNP is likely to be the residues that contribute to NPR-A activation.

7.4 Discussion

[0270] Thus, by in silico conformation analysis, it was speculated that, in human BNP, the amino acid residues that are present in its cyclic part are likely to contribute to NPR-A activation, whereas the amino acid residues present in the tail part are not. It was also suggested that BNP cyclic structure is a smaller molecule than BNP-32 and therefore capable of binding easily and quickly. In fact, a clinical application of a peptide having BNP cyclic structure confirmed that BNP cyclic structure provides faster therapeutic effect than BNP-32. This clinical result is consistent with that of the in silico analysis that BNP cyclic structure contributes to NPR-A activation and can bind to it more easily and quickly than BNP-32. Thus, BNP cyclic structure has a superior therapeutic effect than a general BNP peptide, and therefore is a different substance.

[0271] On the other hand, an NMR analysis revealed that ANP does not take any particular conformation in a solution where its conformation greatly wobbles; this is considered to be similar for BNP. Namely, in human BNP, assuming that the amino acid residues in the cyclic part contribute to its binding to human NPR-A, it is speculated that the amino acid residues in the tail part rather prevent human BNP from entering into the narrow BNP binding site sandwiched between A and B strands of human NPR-A due to its large wobbling. Therefore, it is considered that the cyclic part of human BNP is a relatively smaller molecule than conventionally known human BNP, enabling itself to enter into BNP binding site of NPR-A more easily and quickly. Moreover, it is speculated that the the cyclic part of human BNP has a higher affinity to human NPR-A than BNP-32. This supports the clinical outcomes described herein that the peptide having only the cyclic part of BNP exhibits therapeutic effect faster than BNP-32.

[0272] In order to speculate the effect of BNP cyclic structure (B ring) from non-human species on human Type A receptor, complex models of human NPR-A and BNP rings from pig, bird or rat were generated to surmise interaction. The results suggested that a sufficient effect of the invention can be expected by using BNP ring from non-human animal species (e.g., pigs, birds or rats), as long as it shows an affinity to human NPR-A.

8. Speculating Replaceable Amino Acid Residues in BNP Cyclic Moiety

[0273] Using the constructed model structure of the complex, replaceablity of amino acid residues other than those considered to be involved in the interaction was investigated.

[0274] Specifically, in order to investigate whether the peptide in which amino acid residue that is not assumed to be involved in the interaction has been replaced with another amino acid is capable of binding to NPR-A, a mutant model of BNP was generated to analyze the interaction. Swiss-Pdb viewer was used for modeling.

[0275] Specifically, amino acid residues Gly12, Lys14, Asp16, Ile18, Ser21, Ser22, Gly23, Leu24 and Gly25 in human BNP were targeted for the investigation on their replaceability with other amino acids in terms of following points: [0276] No steric hindrance caused by binding to NPR-A. No interatomic collision observed in the model structure of NPR-A and BNP. [0277] No influence on electrostatic potential on surface. [0278] No large increase in intramolecular energy value (no unnatural angle or twist caused in intermolecular binding). [0279] No non-naturally occurring hydrogen bond formed between the strands of NPR-A and BNP and within BNP strand. [0280] No cavity (cavity, niche) formation.

[0281] Among those described above, intramolecular energy was calculated by ComputeEnergy command of Swiss-Pdb viewer. Intramolecular energy was calculated from the sum of the length of binding, bond angle, twist and binding energy, etc. in the unit kilojoule/mol (Kj/mol).

[0282] The results of analysis indicated the presence of replaceable amino acid residues in Gly12, Lys14, Ile18, Ser21, Ser22, Leu24 and Gly25 among the amino acid residues of human BNP (Table below).

TABLE-US-00028 TABLE 28 human BNP Replaceable amino acid Gly12 Ala, Val, Ser, Thr Lys14 Arg Asp16 none Ile18 Val Ser21 Thr, Ala, Val, Gln, Leu, Ile, Met Ser22 Thr, Ala, Val Gly23 none Leu24 Ala, Val, Ile, Met Gly25 Ala, Ser

[0283] Accordingly, even when the replacement of the amino acid corresponding to those described in the table above took place in the cyclic part of human BNP, i.e., the peptide expressed by the Formula I-a, it was suggested that the peptide replaced in such a way exerts a similar effect as the peptide composed of the amino acid sequence expressed by Formula I-a.

9. Confirming Therapeutic Effect of the Cyclic Peptide with Amino Acid Replacement

[0284] Next, cyclic peptides with some replaced amino acid were subjected to following examination in order to confirm their effect. The methods for preparing the above cyclic peptide and confirming amino acid sequence of the prepared peptide were same as the method for preparing the cyclic peptide as described above and Mass spectroscopy method.

9.1 Confirming Effects on Scalp or Hair

[0285] The obtained cyclic peptide was prepared in purified water at 3, 15, 30, 100 or 500 g/ml and applied to the affected site. Each of formulations A to E in the table indicates the concentration of cyclic peptide at 3, 15, 30, 100 or 500 g/ml, respectively. The absence of the effect upon the application of purified water confirmed the effect was not a placebo effect.

TABLE-US-00029 TABLE29 SEQIDNO (aminoacid For- sequence)of mu- thecyclic la- Diagnosticimpressionofhair peptideused ID Sex Age Cases tion Treatment andscalpafterapplication SEQIDNO16: S1 male 50's multiplealopecia C onceaday After1day,fluffyhairs (CFVRKMDRISS areata startedtogrow.After3days, SSGLGC) obviouslyHairstartedtogrow, restorationandelongationwere observed.Hairgainedelasticity andresilience,thickened.Hair rootregenerationconfirmed. S2 fe- 40's seborrheicderma- C onceaday Icheswereremittedimmediately male titismaleAGA afterapplicaation.After3 minutes,seborrheaanderythema remitted.After1day,dandruff, rednessandseborrheaofscalp improved,fallinghairsreduced. After7days,hairsstimulated togrow,restoredhairandhair growthconfirmed. S3 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume. S4 fe- 50's hairthinning C onceaday After5days,hairstimulation male afteranticancer andgrowthconfirmed.Hairbecame agenttherapy denseandthick. S5 fe- 20's multiplealopecia C onceaday Icheswereimprovedimmediately male areata afterapplication.After1day, fluffyhairsstartedtogrow. After3days,obviouslyHair growthstimulatedandrestored elongationofhairobserved,hair gainedelasticityandresilience, thickened.Hairrootregeneration observed. S6 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO17: S7 male 40's multiplealopecia B onceaday After3days,remarkableterminal (CFGQKMDRISSS areata hairgrowthwasobserved. SGLGC) S8 fe- 20's multiplealopecia B onceaday After5days,newlybornhair male areata rootsandgrownhairswere observed. S9 male 30's maleAGA B onceaday After2or3days,hairgained elasticityandresilience.After 7days,hairgainedvolume, restorationwasconfirmed. S10 fe- 30's femalealopecia C onceaday Hairgainedelasticityandresi- male lience,hairvolumewasincreased onthenightoffirstapplica- tion.After5days,appearanceof scalpthroughhairwereimproved. Theeffectlastedfor1week afterstoppingapplication. SEQIDNO18: S11 male 60's malealopecia B onceaday After1week,hairgainedthick- (CFGHKMDRISSS nessandelasticity,hairdensity SGLGC) wasincreased. S12 male 70's healthyskin C onceaday After1day,hairbecamefuller (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. S13 fe- 40's alopeciaareata B onceaday After5days,hairstartedto male grow,elongationandgrowthof hairwereobserved. S14 fe- 60's alopeciaareata B onceaday After5days,hairstartedto male grow,hairdensitywasincreased. SEQIDNO19: S15 fe- 20's multiplealopecia B onceaday After5days,hairstartedto (CFGRRMDRISSS male areata grow,elongationandrestoration SGLGC) ofhairwereobserved. S16 fe- 50's hairthinning C onceaday After2days,hairgainedelasti- male afteranticancer cityandresiliencevolumeof agenttherapy hairincreased S17 male 60's maleAGA C onceaday After1day,hairgainedelasti- city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed S18 fe- 40's femalealopecia D onceaday After3days,scalpshownthrough male hairduetothinninggotob- secure.Reducedfallinghairs. S19 fe- 40's scalpseborrheic C onceaday After3days,seborrheaand male dermatitis erythemawereimproved,sowas dandruff. SEQIDNO20: S20 male 50's maleAGA C onceaday After1day,hairgainedelasti- (CFGRKLDRISSS city,resilienceandincreased SGLGC) volume. S21 male 40's multiplealopecia C onceaday After3days,hairdensitywas areata increased.Hairstartedtogrow andgrew. SEQIDNO21: S22 male 40's multiplealopecia C onceaday After3days,hairstartedto (CFGRKIDRISSS areata growatthesiteofalopecia SGLGC) areataandgrew. S23 male 40's malealopecia C 1applica- After3days,hairstartedto tion/dayon gainresilience,feltthick. parietal After7days,parietalthinning malethin- wereimprovedandhairvolume ningarea startedtoincrease. S24 fe- 40's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S25 fe- 30's multiplealopecia C onceaday After1day,hairstartedto male areata grow.After2weeks,obviouslots ofhairsstartedtogrow,restor- ation,elongationandgrowthwere observed. S26 fe- 10's multipleophiasis C onceaday After2days,stimulatedhair male growthwasconfirmed.After7 days,obviousstimulatedhair growthwasobserved,elongated andgrownhairwasalsoobserved. SEQIDNO22: S27 male 40's maleAGA B 1applica- After2or3days,hairgained (CFGRKMDRVSSS tion/dayon elasticityandresilience.After SGLGC) forehead 7days,hairgainedvolume, thinning restorationwasconfirmed. area S28 male 20's multiplealopecia C onceaday After3days,hairstartedgrow areata andgrew. S29 fe- 60's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume. S30 fe- 70's femalealopecia C onceaday After7days,increasedhair male volume.Appearanceofscalpshown throughhairgotobsecure. S31 fe- 20's ophiasis B onceaday After5days,stimulatedhair male confirmedalongintractablehair- line,andelongationofhair observed. S32 fe- 30's multiplealopecia C onceaday After1day,hairstartedto male areata grow.After6weeks,obviousstim- ulatedhairgrowth,elongation andgrowthofhairwereobserved SEQIDNO23: S33 male 30's alopeciaareata C onceaday After3days,stimulatedand (CFGRKMDRIGSS universalis elongatedhairswereobserved SGLGC) S34 fe- 60's hairthinning C onceaday After1day,hairgainedelasti- male afteranticancer cityandresilience. agenttherapy S35 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S36 fe- 40's femalealopecia B 1applica- After5days,hairgainedelasti- male tion/dayon city,resilienceandincreased parietal volume.Appearanceofscalp andfore- shownthroughhairgotobsecure. headthin- ningarea S37 male 60's multiplealopecia C onceaday After1day,stimulatedhairwas areata observed.After3days,applica- tionwasstoped,butstimulation andelongationofhaircontined. Whitehairsturnedblackagain, blackhairsstartedtogrow. S38 fe- 40's femalealopecia B onceaday After7days,hairgainedvolume, male seborrheicderma- appearanceofscalpshownthrough titis hairgotobsecure.Seborrheaof scalpwasimproved. S39 fe- 10's ophiasis B onceaday After4days,regardlessofhigh- male lyintractableophiasis,stimu- latedsofthairswereobserved. SEQIDNO24: S40 fe- 10's alopeciaareata A onceaday After7days,stimulatedand (CFGRKMDRISAS male grownsofthairswereobserved. SGLGC) S41 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume. S42 male 30's multiplealopecia B onceaday After4days,stimulationand areata elongationofhairobserved. S43 fe- 30's multiplealopecia C onceaday After1day,hairstartedto male areata grow.Anumberofstimulated, elongatedandgrownhairswere observedin3weeks. SEQIDNO25: S44 male 60's multiplealopecia B onceaday After5days,hairsstartedto (CFGRKMDRISSQ areata growandgrew. SGLGC) S45 fe- 50's maleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S46 fe- 60's multiplealopecia B onceaday After5days,hairrootregenera- male areata tionandhairgrowthstimulation wereobserved.Applicationcon- tinuedandaremarkablerestora- tion,elongationandgrowthof hairwereobserved.Hairelonga- tionratewasfast. S47 fe- 40's femalealopecia C onceaday After1day,hairgainedelasti- male cityandresilience.Application continuedfor3daysandstopped, buthairkeptelasticityand resilience.Theeffectlastedfor 1week. SEQIDNO26: S48 fe- 30's femalealopecia C onceaday After3days,appearanceofscalp (CFGRKMDRISSV male shownthroughhairgotobsecure, SGLGC) hairgainedelasticity,resili- enceandincreasedvolume.Hairs wererestored. S49 male 30's malealopecia B onceaday After5days,volumeofhair increased.Hairgainedelasticity andresilience. S50 fe- 20's femaleAGA C onceaday Volumeofhairincreasedand male scalpdoesnotshowthroughafter 3days.Theeffectwenton S51 fe- 60's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S52 male 20's multiplealopecia C onceaday After3days,stimulatedhairwas areata observed After7days,furtherstimulation andgrowthofhairwerecon- firmed. S53 fe- 30's multiplealopecia C onceaday After1day,hairstartedto male areata grow.Terminalhairsstartedto grow,andrestoration,elongation andgrowthofhairwereobserved in2weeks. SEQIDNO27: S54 fe- 50's hairthinning B onceaday After5days,hairgotthickand (CFGRKMDRISSL male afteranticancer dense. SGLGC) agenttherapy S55 male 50's maleAGA C onceaday After1day,hairgainedelasti- city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S56 male 20's multiplealopecia E onceaday After3days,hairstartedto areata grow After7days,stimulationand growthofhairwereconfirmed. SEQIDNO28: S57 fe- 40's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISSI male city,resilienceandincreased SGLGC) volume. S58 male 20's multiplealopecia C onceaday After1day,hairfallingwasre- areata ducedandhairstartedtogrow. After7days,remarkablestimula- tion,elongationandgrowthof hairwereconfirmed. S59 male 20's multiplealopecia C onceaday After1day,hairstartedtogrow areata andfallinghairsreduced.After 4days,applicationwasstoped, butstimulation,elongationand restorationofhairpersisted. S60 fe- 50's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. S61 fe- 40's multiplealopecia C onceaday After3days,hairsstartedto male areata growandgrew. S62 fe- 40's alopeciaareata B onceaday After5days,stimulatedhair male growthwasconfirmed.After2 weeks,furthergrowthofterminal hair,elongationandgrowthof hairwereconfirmedallover alopeciasite. SEQIDNO29: S63 fe- 40's femalealopecia C onceaday After4days,hairgainedelasti- (CFGRKMDRISSM male city,resilienceandincreased SGLGC) volume. S64 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. S65 male 40's maleAGA C onceaday Hairgainedresilienceandvolume after3days.Applicationcon- tinuedfor1week,andtheeffect persistedfor1.5monthsthere- after. S66 male 30's malealopecia C 2applica- After3days,hairgainedelasti- tion/day city,resilienceandincreased for3days volumeatthinningareaonfore- head.Afterafewdays,hair growthstimulationandhair growthwereconfirmed. S67 fe- 70's femalealopecia C onceaday After2days,hairgainedelasti- male cityandresilience.After5 days,volumeofhairincreased. Itwasconfirmedthatthestimu- latedhairsarepredominantly blackoverwhitehair. SEQIDNO30: S68 fe- 20's alopeciaareata B onceaday After7days,stimulatedhair (CFGRKMDRISSS male ophiasis growthwasconfirmed.Even2 VGLGC) weeksafterstoppingapplication, hairgrowthstimulationandre- storationpersisted,andthere werelittlefallinghairs. S69 male 40's maleAGA B onceaday After2-3days,hairgained elasticityandresilience.After 7days,hairgainedvolume,re- storationwasconfirmed. S70 male 30's alopeciauniver- B onceaday After3days,hairsstartedto salis growandgrew. S71 fe- 30's multiplealopecia C onceaday After1day,stimulatedhair male areata growthwasconfirmed. SEQIDNO31: S72 fe- 20's alopeciaareata C onceaday After7days,stimulatedhair (CFGRKMDRISSS male ophiasis growthwasconfirmed.Even2 SRLGC) weeksafterstoppingapplication, hairgrowthstimulationandre- storationpersisted,andfalling hairssignificantlydecreased S73 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. S74 male 30's alopeciaareata C 2applica- After3days,stimulationand universalis tion/day elongationofhairwerecon- for3days firmed. S75 fe- 60's multiplealopecia B onceaday After5days,hairsstimulatedto male grow,restoredhairandhair growthwereconfirmed.Itwas confirmedthathairelongation rateisfast. SEQIDNO32: S76 male 30's malealopecia B onceaday After7days,hairgainedelasti- (CFGRKMDRISSS city.Hairstartedtogrow,and SGMGC) volumeofhaironparietalfore- headwasincreased. S77 fe- 60's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S78 male 40's alopeciauniver- C onceaday After3days,hairstimulation salis andgrowthwereconfirmed S79 fe- 50's hairthinning C onceaday After2days,hairgainedelasti- male afteranticancer city,resilienceandincreased agenttherapy volume.Appearanceofscalpshown throughhairgotobsecure.Hair increasewasconfirmed. SEQIDNO33: S80 fe- 50's multiplealopecia B onceaday After7days,stimulatedhair (CFGRKMDRISSS male areata growthwasconfirmed. SGIGC) S81 male 60's healthyskin C onceaday After1day,hairbecamefuller (scalp withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. S82 fe- 40's multiplealopecia B onceaday After5days,hairrootregenera- male areata tionandhairgrowthstimulation wereconfirmed. S83 male 60's alopeciaareata C onceaday After1day,apparenthairgrowth stimulationandrestorationwere confirmed.After4days,hair elongationwereconfirmed. SEQIDNO34: S84 fe- 50's hairthinning B onceaday After5days,thin,fuzzyhairs (CFGRKMDRISSS male afteranticancer alonghairlinegotthickerand SGVGC) agenttherapy denser. S85 male 50's maleAGA C onceaday After1day,hairgainedelasti- city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S86 male 40's maleAGA C onceaday Hairgainedresilienceandvolume after3days.Applicationcon- tinuedfor1week,andtheeffect persistedfor1.5monthsthere- after. S87 male 30's malealopecia C onceaday After3days,hairgainedresili- enceandvolumeofhairincreased inparietalthinningarea S88 male 20's multiplealopecia C onceaday After1day,hairsstartedto areata growandgrew. SEQIDNO35: S89 fe- 40's hairthinning B onceaday After5days,hairgainedelasti- (CFGRKMDRISSS male afteranticancer city,resilience,thickness,and SGAGC) agenttherapy increasedvolume. S90 male 20's multiplealopecia C onceaday After3days,stimulatedhair areata growthwasconfirmed. S91 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. S92 fe- 60's multiplealopecia B onceaday After5days,hairrootregenera- male areata tionandhairgrowthstimulation wereobserved.Then,remarkable restoration,elongationand growthofhairwereobserved. S93 fe- 30's multiplealopecia C onceaday After1day,stimulatedhairwas male areata observed.After2weeks,thereis remarkableelongationofhair marginalareaaroundlargede- calvantspot,andnewhairstart- edtogrowincenterpart. SEQIDNO36: S94 male 30's alopeciauniver- C onceaday After4days,hairstimulation (CFGRKMDRISSS salis andgrowthwereconfirmed. SGLSC) S95 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. S96 male 50's seborrheicderma- C onceaday After3days,seborrheaand titis erythemawereimproved. S97 male 50's malealopecia B onceaday After5days,hairgainedelasti- city,resilienceandincreased volume. SEQIDNO37: S98 fe- 40's hairthinning B onceaday After5days,hairgainedelasti- (CFGRKMDRISSS male afteranticancer city,resilience,thicknessand SGLAC) agenttherapy increasedvolume. S99 male 40's maleAGA B 1applica- After2or3days,hairgained tion/dayon elasticityandresilience.After forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S100 male 20's multiplealopecia C onceaday After5days,stimulatedhair areata growthwasconfirmed. S101 fe- 20's alopeciaareata A onceaday After10days,hairgrowthstimu- male lationelongationwereconfirmed. SEQIDNO38: S102 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- (CFARKMDRISSS male city,resilienceandincreased SGLGC) volume.Hairstartedtogrow, restorationandelongationwere observed. S103 fe- 50's dandruff,itches C onceaday Ichesstoppedin3minutes, male dandruffwasreduced,rednessin scalpwassignificantlyimproved. S104 fe- 10's eclopicmultple C onceaday After3days,stimulatedhair male alopecia growthwasconfirmedineyebrows. SEQIDNO39: S105 male 10's alopeciauniver- C onceaday After3days,stimulatedhair (CFSRKMDRISSS salis growthwasconfirmed. SGLGC) S106 fe- 50's dandruff,itches C onceaday Ichesstoppedin1minute,dan- male druffwasreduced,rednessin scalpwassignificantlyimproved. S107 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. S108 fe- 60's multiplealopecia C onceaday After3days,stimulatedhair male areata growthwasconfirmed.Moreblack hairsstartedtogrowthanwhite hairs,andhairelongationrate wasfast. SEQIDNO40: S109 male 60's maleAGA C onceaday After1day,hairgainedelasti- (CFTRKMDRISSS city,resilienceandincreased SGLGC) volume.Hairstartedtogrow, restorationandelongationwere observed. S110 male 20's multiplealopecia C onceaday After4days,stimulatedhair areata growthwasconfirmed C onceaday SEQIDNO41: S111 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISST male city,resilienceandincreased SGLGC) volume.Hairstartedtogrow, retorationandelongationwere observed. S112 male 20's multiplealopecia C onceaday After4days,stimulatedhair areata growthwasconfirmed. SEQIDNO42: S113 male 30's multiplealopecia C onceaday After3days,stimulatedhair (CFGRKMDRISSA areata growthwasconfirmed. SGLGC) S114 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. S115 fe- 60's multiplealopecia C onceaday After3days,stimulatedhair male areata growthwasconfirmed.Blackhairs startedtogrowandhairelonga- tionwasfast SEQIDNO43: S116 male 40's maleAGA B 1applica- After2-3days,hairgained (CFGRKMDRISSS tion/dayon elasticityandresilience.After TGLGC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed area S117 male 20's multiplealopecia D onceaday After3days,stimulatedhair areata growthwasconfirmed. SEQIDNO44: S118 fe- 60's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISSS male city,resilienceandincreased AGLGC) volume.Hairstartedtogrow, restorationandelongationwere observed S119 fe- 20's multiplealopecia C onceaday After3days,stimulatedhair male areata growthwasconfirmed. SEQIDNO45: S120 male 40's maleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISSS city,resilienceandincreased SGLGC) volume.Hairstartedtogrow, restorationandelongationwere observed. S121 fe- 40's healthyskin C onceaday After1day,hairvolumewasin- male (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO46: S122 fe- 60's femaleAGA C onceaday After1day,hairgainedelasti- (CFSRKMDRISST male city,resilienceandincreased SGLGC) volume.Hairstartedtogrow, restorationandelongationwere observed. S123 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO47: S124 male 40's healthyskin C onceaday After1day,hairvolumewasin- (CFSRKMDRISSS (scalp) creased.Hairgainedelasticity TGLGC) andresilience. S125 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow,re- storationandelongationwere observed. SEQIDNO48: S126 fe- 40's multiplealopecia B onceaday After5days,hairrootregenera- (CFSRKMDRISSS male areata tionandhairgrowthstimulation SGIGC) wereconfirmed S127 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack S128 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow, restorationandelongationwere observed. SEQIDNO49: S129 fe- 60's femaleAGA C onceaday After1day,hairgainedelast- (CFSRKMDRISSS male city,resilienceandincreased SGLAC) volume. S130 fe- 60's alopeciaareata B onceaday After5days,hairstartedto male growandhairdensitywasin- creased S131 fe- 40's healthyskin C onceaday After1day,hairvolumewasin- male (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO50: S132 male 50's maleAGA C onceaday After1day,hairgainedelasti- (CFGRRMDRISST city,resilienceandincreased SGLGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S133 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO51: S134 male 40's maleAGA B 1applica- After2or3days,hairgained (CFGRRMDRISSS tion/dayon elasticityandresilience.After TGLGC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S135 male 50's healthyskin C onceaday After1day,hairbecamefuller (scalp) withincreasedvolume.Hair gainedelasticityandresilience. SEQIDNO52: S136 male 60's maleAGA C onceaday After1day,hairgainedelasti- (CFGRRMDRISSS city,resilienceandincreased SGIGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S137 fe- 40's multiplealopecia B onceaday After5days,hairrootregenera- male areata tionandhairgrowthstimulation wereconfirmed. S138 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO53: S139 fe- 40's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRRMDRISSS male city,resilienceandincreased SGLAC) volume. S140 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO54: S141 male 40's maleAGA B 1applica- After2or3days,hairgained (CFGRKMDRISST tion/dayon elasticityandresilience.After SGIGC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S142 fe- 50's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO55: S143 male 50's maleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISST city,resilienceandincreased SGLAC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S144 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO56: S145 male 40's maleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISSS city,resilienceandincreased TGIGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S146 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO57: S147 male 50's maleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISSS city,resilienceandincreased TGLAC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S148 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience SEQIDNO58: S149 fe- 40's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRKMDRISSS male city,resilienceandincreased SGIAC) volume. S150 fe- 40's healthyskin C onceaday After1day,hairvolumewasin- male (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO59: S151 male 50's maleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISST cityresilienceandincreased SGLGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S152 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO60: S153 male 60's maleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISSS city,resilienceandincreased TGLGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S154 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO61: S155 fe- 60's femaleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISSS male city,resilienceandincreased SGIGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S156 male 60's healthyskin C onceaday After1day,hairbecamefuller (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO62: S157 male 50's maleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISSS city,resilienceandincreased SGLAC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S158 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO63: S159 male 60's maleAGA B 1applica- After2or3days,hairgained (CFSRKMDRISST tion/day elasticityandresilience.After SGIGC) onforehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S160 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO64: S161 male 60's healthyskin C onceaday After1day,hairbecamefuller (CFSRKMDRISSS (scalp) withincreasedvolume.Hair TGIGC) gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. S162 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- male city,resilienceandincreased volume.Hairstartedtogrow,re- storationandelongationwere observed SEQIDNO65: S163 male 40's maleAGA B 1applica- After2or3days,hairgained (CFSRKMDRISSS tion/dayon elasticityandresilience.After TGLAC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S164 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO66: S165 male 60's maleAGA C onceaday After1day,hairgainedelasti- (CFSRKMDRISSS city,resilienceandincreased SGIAC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S166 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO67: S167 male 40's maleAGA B 1applica- After2-3days,hairgained (CFGRRMDRISST tion/dayon elasticityandresilience.After SGIGC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S168 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO68: S169 fe- 40's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRRMDRISST male city,resilienceandincreased SGLAC) volume. S170 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO69: S171 male 60's maleAGA B 1applica- After2or3days,hairgained (CFGRRMDRISSS tion/dayon elasticityandresilience.After TGIGC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S172 male 30's multiplealopecia C onceaday After1day,hairsstartedto areata growandgrew. S173 male 60's healthyskin C onceaday After1day,hairbecamefuller (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO70: S174 fe- 50's femaleAGA C onceaday After1day,hairgainedelasti- (CFGRRMDRISSS male city,resilienceandincreased TGLAC) volume. S175 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO71: S176 male 60's maleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISST city,resilienceandincreased SGIGC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S177 male 20's multiplealopecia C onceaday After1day,hairsstartedto areata growandgrew. S178 male 60's healthyskin C onceaday After1day,hairbecamefuller (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO72: S179 male 40's maleAGA B 1applica- After2or3days,hairgained (CFSRRMDRISST tion/dayon elasticityandresilience.After SGLAC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S180 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO73: S181 fe- 40's femaleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISSS male city,resilienceandincreased TGIGC) volume. S182 fe- 40's alopeciaareata B onceaday After5days,hairstartedto male growandhairdensitywasin- creased. S183 fe- 60's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. SEQIDNO74: S184 male 60's maleAGA C onceaday After1day,hairgainedelasti- (CFSRRMDRISSS city,resilienceandincreased TGLAC) volume.Hairstartedtogrow,re- storationandelongationwere observed. S185 male 40's healthyskin C onceaday After1day,hairvolumewasin- (scalp) creased.Hairgainedelasticity andresilience. SEQIDNO75: S186 male 40's maleAGA B 1applica- After2or3days,hairgained (CFSRRMDRISSS tion/dayon elasticityandresilience.After SGIAC) forehead 7days,hairgainedvolume,re- thinning storationwasconfirmed. area S187 fe- 40's multiplealopecia B onceaday After5days,hairrootregenera- male areata tionandhairgrowthstimulation wereconfirmed. S188 fe- 50's healthyskin C onceaday After1day,hairbecamefuller male (scalp) withincreasedvolume.Hair gainedelasticityandresilience. After7days,grownhairswere predominantlyblackoverwhite hairs.Whitehairsturnedblack. In SEQ ID NOS 16-75. 1st Cys and 17th Cys from left form a disulfide bond.

[0286] The result described above demonstrated that the cyclic peptide of the invention exhibits a similar effect as the cyclic peptide expressed by Formula I-a even if some amino acids of the cyclic peptide have been replaced.

[0287] Specifically, it has effects of preventing hair loss at the applied site, stimulating hair growth or promoting hair growth, giving hair resilience and elasticity, increasing volume of hair and dramatically decreasing falling hairs when it was applied to the site of alopecia or hair growth stimulation, etc. It also has effects of nourishing hair, promoting hair growth stimulation, improving/preventing thinning of hair at the applied site. In this case, the stimulated hair tends to become a terminal (non-white) hair. Moreover, these effect exhibit significantly faster as compared to other active agents conventionally used in alopecia therapeutics such as BNP such that stimulated hair growth can be confirmed one day after only one application even in the case of an intractable alopecia areata multilocularis or ophiasis. In addition, resulting effects persists remarkably even after terminating application and the stimulated hair keep growing. In the case of AGA or healthy subject it exert immediate effect of providing hair with resilience and elasticity on the following day, increasing the volume, and dramatically decreasing falling hairs, which effects last even after terminating application. It also has an improving and preventing effect on dermatitis. Therefore, it can improve or prevent skin inflammation associated with alopecia. Such effect is advantageous in cases where alopecia has been exacerbated due to skin condition of the applied site (e.g., scalp). Moreover, the external preparation of the present invention exerts a moisturizing and skin texture-improving effects at the applied site when being applied to skin as described above. It can remove and suppress dandruff and itching, while giving moisture to hair and scalp, improving and preventing dryness and keeping hair and scalp healthy. It also can improve seborrhea. On the other hand, when it is used as an alopecia therapeutic/prophylactic, it can be used for the purpose of treating or preventing one or more of applicable alopecia including, for example, those described below, without being particularly limited thereto.

(Acquired Alopecia)

[0288] (i) Alopecia without accompanying scarring or skin lesion (alopecia areata, male pattern alopecia, seborrheic alopecia, alopecia pityroides, female pattern alopecia, gestational alopecia, malignant alopecia, senile alopecia, alopecia totalis, alopecia areata multilocularis, ophiasis, drug-induced alopecia, cancer chemotherapy-induced alopecia and radiation exposure-induced alopecia, traumatic/mechanical alopecia, malnutrition/metabolic disorder associated alopecia, endocrine dysfunction associated alopecia and telogen effluvium (post partum alopecia, alopecia after high fever)).
(ii) Alopecia observed in skin lesion or pathologic skin (infection-induced alopecia, tumor-induced alopecia, inflammation-induced alopecia).
(iii) Scarring alopecia (skin infection-induced alopecia, alopecia induced by infiltration of inflammatory cells).

(Congenital Alopecia)

[0289] Diffuse alopecia, congenital atrichia, alopecia in hereditary syndromes, localized alopecia, phakomatosis, aplasia cutis, congenital alopecia triangularis.

[0290] It exerts an excellent effect especially on acquired alopecia, preferably on alopecia without accompanying scarring or skin lesion, more preferably on alopecia areata, male pattern alopecia, seborrheic alopecia, alopecia pityroides, female pattern alopecia, gestational alopecia, malignant alopecia, senile alopecia, alopecia totalis, alopecia areata multilocularis, ophiasis, drug-induced alopecia, cancer chemotherapy-induced alopecia and radiation exposure-induced alopecia, traumatic/mechanical alopecia, malnutrition/metabolic disorder associated alopecia, endocrine dysfunction associated alopecia and telogen effluvium. Therefore, the external preparation of the present invention can be used for the purpose of treating or preventing at least one or more alopecia selected from the above-mentioned group.

[0291] One simple application onto aforementioned affected site at 3 g/ml to 500 g/ml improved or eliminated itch, dandruff, scalp redness and inflammation immediately thereafter (immediate effect). A single application stimulated hair growth, grew hair, provided hair with resilience, elasticity and increased volume, and dramatically decreased falling hairs, and these effects started on the day after the application and persisted for one week or more (long-lasting effect). If application was carried on for about one week, these effects are further improved, and even after terminating application, the effects lasted from 1 to 2 weeks such that stimulation of hair growth and restoration of hair continued and there were little or no falling hairs.

9.2 Confirming Effects on Skin

[0292] The obtained cyclic peptide was prepared in purified water at 3, 15, 30, 100 or 500 g/ml and applied to affected site. Each of formulations A to E in the table indicates the concentration of the cyclic peptide at 3, 15, 30, 100 or 500 g/ml, respectively. The absence of the effect upon the application of purified water without the peptide confirmed the effect was not a placebo effect.

TABLE-US-00030 TABLE30 Aminoacidsequence ofcyclicpeptide Diagnostic Diagnosticimpressiononskin Itchesafter mutantused ID Sex Age impression Formulation Treatment afterapplication application 1 SEQIDNO16: Q1 female 30s eczema B onceaday After3minutes,redpapulewasreduced Itcheswereremitted (CFVRKMDRISSSSGLGC) andremitted. immediatelyafter application. Q2 male 20s atopic B onceaday After2minutes,erythema,edemaand Itcheswereeliminated dermatitis infiltrationwasremitted. immediatelyafter application. Q3 male 80s plaque C onceaday,2 After3minutes,crusts,scales,erythema psoriasis days andinfiltrationwereremitted. Q4 male 40s plaque C onceaday, After10minutes,scales,erythemaand psoriasis infiltrationwereremitted. Q5 female 30s eczema C onceaday After3minutes,erythemawasremitted. Q6 male 20s atopic B onceaday.7 Erythema,infiltrationandpapulewere Itcheswereremitted dermatitis days remitted. immediatelyafter application. Q7 female 60s eczema C onceaday After2minutes,erythema,infiltration andpapulewereremitted. After20minutes,theywereamaliorated. Q8 female 20s atopic C onceaday After2minutes,erythemaand Itcheswereeliminated dermatitis infiltrationwereremitted. in1minute Q9 male 10s atopic C onceaday Rednessvanishedimmediatelyafter Itcheswereextincted dermatitis application. in1minute. SEQIDNO17: Q10 male 40s atopic C onceaday After3minutes,erythemaand After3minutes, (CFGQKMDRISSSSGLGC) dermatitis infiltrationwereremitted. itcheswereremitted. Q11 male 10s nummular C onceaday After3minutes,infiltration,erythema eczema exudateandscaleswereremitted. Q12 male 20s eczema B onceaday After2minutes,scalesanderythema Icheswereelimintaed wereremitted.Withonly1application, immediatelyafter norecurrencefor7daysthereafter. application. 2 SEQIDNO18: Q13 male 60s milleria C onceaday After2minutes,rednesswasremitted. (CFGQKMDRISSSSGLGC) Q14 female 40s roughened A onceaday After2minutes,zits,drynessand skin rawnessonskinwerereduced,skinwas moisturizedandittextureimproved. After7days,largewrinklesand nasolabialfoldbecamelessdeep. SEQIDNO19: Q15 male 20s eczema,atopic C onceaday After2minutes,erythemaandscaleswere (CFGRRMDRISSSSGLGC) dermatitis remitted.After20minutes,skingot moisturized.Afteronly1application, erythemavanishesandkeptinagood conditionfor1week. Q16 male 20s atopic C onceaday After5minutes,woundwasapithelized, dermatitis erythemaandpapulewereimproved. Q17 female 40s roughened B onceaday After2minutes,rednesswasremitted. skin Roughnessandskintexturewerealso improved. Q18 female 30s roughened B onceaday After2minutes,wasremitted.Roughness skin andskintexturewerealsoimproved. Rawnessgotbetter,ichesvanished immediatelyafterapplication,skin weremoisturizedandsmooth. Q19 male 40s plaque C onceaday After3minutes,erythema,crustsscales psoriasis andinfiltrationwereremitted. Q20 male 40s eczema C onceaday Erythemawasremittedimmediatelyafter Itcheswereextincted application. immediatelyafter application SEQIDNO20: Q21 female 60s eczema C onceaday After1minute,redpapulewasreduced After3minutes, (CFGRKLDRISSSSGLGC) andextincted. icheswereremitted Q22 male 20s atopic B onceaday After2minutes,erythemaandscales Itchesvanished dermatitis wereremitted. completelyimmediately afterapplication. Q23 female 40s eczema C onceaday After2minutes,erythema,scalesand Rawnessvanished papulewereremitted. immediatelyafter Q24 female 30s roughened B onceaday After3minutes,zits,ruggedness,dry Ichesapplicationwere skin,acne skinandrednesswereremitted,skingot eliminatedimmediately fullanditstextureimproved.Acnes afterapplication. werereducedandrednesswasremitted. Q25 female 60s wrinkles, B onceaday After2minutes,wrinklesbecameless dryskin deep,dryskinwasremitted. Q26 female 60s eczema C onceaday After3minutes,erythemaandscales wereremitted. SEQIDNO21: Q27 male 20s eczema,atopic C onceaday After3days,erythema,scalesand Icheswereeliminated (CFGQKIDRISSSSGLGC) dermatitis papulewereimproved.Crackswere immediatelyafter epithelized,skingotmoisturizedand application. improved. Q28 female 40s eczema B onceaday After2minutes,infiltration,erythema, After1minute,iches papuleandscratchwoundwereremitted. wereeliminated. Q29 male 40s atopic B onceaday Immediatelyafterapplication,erythema After3minutes, dermatitis andscalesandremitted. icheswereeliminated. 3 SEQIDNO22: Q30 female 30s atopic C onceaday After3minutes,1application,erythema After30seconds, (CFGQKMDRISSSSGLGC) dermatitis andinfiltrationwereremarkablyremitted, Icheswereeliminated. dryskin,scalesandcrackswerealso improved. Q31 female 70s eczema C onceaday immediatelyafterapplication,erythema Facialrawness andpapuleremitted. vanishedimmediately Q32 female 30s atopic B onceaday, Aremarkableimprovementafter2days, afterapplication. dermatitis 7days anderythemawaseliminated. Q33 female 40s atopic C onceaday, After1minute,erythemawasremitted. Icheswereeliminated dermatitis 2days After2minutes,scratchwoundwas immediatelyafter epithelized. application After3minutes,skintexturewasimproved. Icheswereeliminated Withonly1-dayapplicatioin,erythema immediatelyafter waseliminated,skintexturewas application improvedwithnodryness. Q34 male 20s atopic C onceaday After2minutes,erythemaandscaleswere Icheswereremitted dermatitis remitted.Norecurrencefor7days immediately thereafter. afterapplication Q35 female 50s chroniceczema C onceaday After2applications,lichenification Icheswereeliminated erythemawasremitted.Norecurrence immediatelyafter for3weeksthereafter. application Q36 female 50s roughened C onceaday Skinroughnesswascuredimmediately skin afterapplication.After2minutes,skin gotmoisturized.Itsrednessfaded.Skin texturewasfinelyimproved.After7 days,skintonebecamebrighter,spots fadedandflabinesswasimproved. Q37 female 30s roughened B onceaday After2minutes,zits,rednesswas Rawnessvanished skin remitted.After1day,skinbecame immediatelyafter normaltoneanditstexturewasimproved. application. SEQIDNO23: Q38 female 40s atopic C onceaday After2minutes,scratchwoundwas After1minute,acnes (CFGQKMDRISSSSGLGC) dermatitis epithelizedandcured.Erythemawas wereeliminated. improved. Q39 male 20s eczema C onceaday After2minutes,erythema,dryskinand Itcheswereabeled scaleswereimprovedandskingot immediatelyafter moisturized. application Q40 male 20s atopic C onceaday After2minutes,erythema,scalesand Itcheswereextincted dermatitis infiltrationwereremitted.With1 immediatelyafter application,norecurrencefor7days. application. Q41 male 40s plaque C onceaday After3minutes,crustsscaleserythema psoriasis andinfiltrationwereremitted. Q42 female 30s roughened B onceaday After2minutes,roughness,zitsand After2minutes, skin rednesswerereduced,skingotmoisturized itcheswere anditstexturewasimproved. extincted. Q43 female 50s rosacea,acne C onceaday After1minute,erythema,telangiectasia andacnewereremitted.After2minutes, papule,infiltration,acne,dryskinand rednesswasremitted. 4 SEQIDNO24: Q44 male 10s atopic C onceaday After3minutes,erythema,scalesand (CFGQKMDRISASSGLGC) dermatitis infiltrationwereremitted. Furtherimprovementafter40minutes. Scratchwoundwasepithelized. q45 male 20s atopic C onceaday After3minutes,erythemaand After3minutes, dermatitis infiltrationwereremitted. Icheswere Q46 male 20s C onceaday After3days,crackswereepithelized. eliminated. Erythema,scalesandinfiltrationwere remitted. Q47 male 20s atopic C onceaday After2minutes,woundwasepithelized. Icheswereeliminated dermatitis Erythema,scalesandlicherification immediatelyafter remitted.After5minutes,skingot application. softened,erythemaandinfiltrationwere improved. SEQIDNO25: Q48 male 20s C onceaday After2minutes,erythema,scalesand Icheswereremitted (CFGQKMDRISSQSGLGC) infiltrationwereremitted.Skingot immediatelyafter moisturized. application. Q49 female 40s roughened B onceaday After3minutes,deepwrinkleswereless Rawnesswaseliminated skin deep,dryskinandrednesswereimproved, immediatelyafter skingotmoisturizedandfull.Byusing application. for1week,largewrinklesandnasolabial foldbecamelessdeep.Skinbecame brighter.Spotsfaded.Flabinesswas improved. SEQIDNO26: Q50 male 60s plaque C onceaday After2minutes,scalesandinfiltration (CFGQKMDRISSSSGLGC) psoriasis remitted. Q51 male 30s millaria C onceaday After2minutes,remitted. Q52 male 20s acnevulgaris C onceaday After6minutes,acnebecamedryandwas reducedinsize. Q53 female 60s eczema C onceaday After3minutes,erythemaandinfiltration wereremitted. After4minutes,amelicrated. Q54 female 10s atopic C onceaday After2minutes,strongitchesalmost dermatitis vanished.Erythemaandlichenification wereimproved.Skingotsoftened. Q55 female 40s roughened C onceaday After2minutes,zits,rednessand Rawnessvanished skin roughnesswereimproved.Skingot immediatelyafter moisturizedanditstexturewas application. improved.Skinwasimproved. SEQIDNO27: Q56 female 50s roughened C onceaday After3minutes,dryskinwasimproved, (CFGQKMDRISSSSGLGC) skin,acne skingotmoisturizedanditstexturewas finelyimproved.Rednessvanished. Inframmationinacnewasremitted, rednessfadedanddried.Byusingfor1 week,skinbecamebrighter. Q57 male 20s atopic C onceaday After3minutes,erythema,infiltration Icheswereeliminated dermatitis andlichenificationwasimproved. immediatelyafter application. 5 SEQIDNO28: Q58 female 30s eczema C onceaday After1minute,erythemaandinfiltration After1minute, (CFGQKMDRISSISGLGC) wereremitted. Icheswere Q59 female 20s roughened A onceaday After7-daycontinuousapplication,skin eliminated. skin roughnessvanished,wrinkleswereless deep,andskinbecamebrighter. Q60 female 20s roughened C onceaday After2minutes,scaleswereimproved. skin With1application/day,roughnesswas improvedandskincouldbekept moisturized. SEQIDNO29: Q61 female 30s eczema C onceaday After1minute,erythemaand Icheswereeliminated (CFGQKMDRISSMSGLGC) infiltrationwereremitted. immediatelyafter application. SEQIDNO30: Q62 male 20s atopic B onceaday, After3days,erythemaandinfiltration (CFGQKMDRISSSVGLGC) dermatitis 7days wereremitted. Q63 female 60s eczema C onceaday After1minute,erythemaandpapulewere Icheswereeliminated remitted.After3days,therewere immediatelyafter almostcured. application. Q64 male 50s dryskin C onceaday Immediatelyafterapplication,skingot promptlymoisturizedanditstexturewas improved,zits,erythemaandroughned skinwereimproved. Q65 male 20s atopic C onceaday After10minutes,lichenificationwas Icheswereeliminated dermatitis improved.Skingotsoftened,erythema immediatelyafter andscaleswereimproved. application. Q66 female 40s roughened A onceaday After1daywith1application,skin skin conditionsofroughness,zitsanditches wereimprovedandskintexturewasfinely improved.Theeffectlastedfor7days thereafter. SEQIDNO31: Q67 male 20s atopic C onceaday After2minutes,erythemaandpapule (CFGQKMDRISSSSRLGC) dermatitis wasremitted. SEQIDNO32: Q68 male 30s millaria C onceaday After2minutes,redpapulewasreduced (CFGQKMDRISSSSGMGC) insize,skindrynesswasremitted. Q69 male 10s atopic C onceaday After3minutes,exudatestopped, dermatitis, erythemaandinfiltrationwereimproved. numeuler 6 SEQIDNO33: Q70 female 30s contact C 2applicatioin After3minuts,erythema,infiltration After1minute,iches (CFGQKMDRISSSSGIGC) dermatits andeczemawereremitted. wereeliminated. Q71 female 30s atopic E onceaday Immediatelyafterapplication,erythema Icheswereeliminated dermatitis andscaleswereimproved. immediatelyafter Q72 female 70s plaque C 2application After5minutes.erythema,scales application. psoriasis andinfiltrationwereremitted. Q73 female 40s roughened B onceaday In2minutesafterapplication,skin skin wasimproved.Skintexturewasfinely improved.Skingotsoftened. SEQIDNO34: Q74 female 30s eczema C 2applicatioin After3minutes,erythema,infiltration After1minute,Iches (CFGQKMDRISSSSGVGC) andedemaremitted. wereeliminated. Q75 male 20s eczema,acne C 2applicatioin After5minutes,acnewasdriedand vulgaris reducedinsize.Erythemaandscales wereremitted. Q76 female 70s plaque C 2applicatioin After5minutes,erythema,scalesand psoriasis infiltrationwereremitted. Q77 female 40s roughened C onceaday After2minutes,skintexturewas skin improved,skingothull,andredness anddrynesswereimproved. Q78 male 50s roughened B onceaday After3minutes,skingotmoisturized skin anditstexturewasimproved. Q79 female 20s atopic C onceaday After3minutes,erythemaandscales Itcheswereimproved dermatitis wereimproved. immediatelyafter applilcation. Q80 female 50s roughened B onceaday After3minutes,skinroughnesswas Ichesweeelminated skin improved,skingotmoisturizedandits immediatelyafter texturewasfimelyimproved.Skingained application. resilence.After7days,largewrinkles andnasolabialfoldbecauselessdeep. Bycontinuing4weeks,spotsfadedand skintonebecamebrighter.Skinresilence wasimproved.Largewrinklesbecameless deep. Q81 male 30s roughened B onceaday After3minutes,skinroughnessand skin rednesswereimproved.Skintexture wasfinelyimproved.Bycontinuingfor 7days,skinresilencewasimproved. SEQIDNO35: Q82 female 40s roughened B onceaday After3minutes,roughnessskinanddry (CFGQKMDRISSSSGAGC) skin skinwerepromptlyimproved,skin texturewasimproved. Q83 male 60s plaque C twiceaday After30minutes,erythema,scalesand psoriasis infiltrationwereimproved. Q84 male 40s plaque C twiceaday After20minutes,erythemaandscales psoriasis wereimproved. Q85 female 70s plaque C twiceaday After5minutes,erythemaandscales psoriasis wereremitted. 7 SEQIDNO36: Q86 female 10s atopic C onceaday After2minutes,linchenificationwas After2minutes, (CFGQKMDRISSSSGLSC) dermatitis remarkablyremitted,erythema,scales strongicheswere inflimation,cracks,scratchwound, eliminated. etc.allremitted.After5minutes, improvedwithonlymildinfiltration left. Q87 female 20s atopic B onceaday After1minute,erythema,infiltration After2minutes, dermatitis andpapuleremitted.After4minutes, icheswere infiltrationwasremittedwithonly eliminated. milderythemaleft. Q88 male 10s atopic C onceaday After3minutes,erythema,scales, After3minutes, dermatitis infiltrationandscratchwoundwere icheswere remitted. eliminated. Q89 female 40s roughened B onceaday Immediatelyafterapplication,itches skin andrednesswereimproved.After3 minutes,skingotfullandresilent, andskinwasimproved. Q90 male 50s seborrhea C onceaday, Immediatelyafterapplication,seborrhea Itcheswereremitted dermatits 3days wasremitted.After3days,erythema immediatelyafter andseborrheawasremiited. application. SEQIDNO37: Q91 male 60s plaque C onceaday After2minutes,erythemaandscales (CFGQKMDRISSSSGLAC) psoriasis wereremitted. Q92 male 20s atopic C onceaday After2minutes,erythemaand Icheswereeliminated dermatitis infiltrationwereremitted.Applied immediatelyafter prurigo twice.After15minutes,pringenodularis application. nodularis, onhipwasremitted Q93 female 40s roughened B onceaday After3minutes,roughenedskinanddry skin skinwereimprovedpromptlyandskin texturewasimproved. Q94 male 40s plaque C onceaday After20minutes,erythemaandscales psoriasis wereremitted. Q95 male 30s atopic C onceaday After2minutes,scratchwound dermatitis epithelizederythemawasremitted. SEQIDNO38: Q96 male 20s atopic C onceaday After3minutes,crackswasreducedin immediatelyafter (CFGQKMDRISSSSGLGC) dermatitis size.Erythema,scalesand application,itches lichenificationwereremitted. andrawnesswere removed. Q97 female 40s roughened B onceaday After1minute,dryskinwasimproved, immediatelyafter skin skingotmoiturizedandfull,and application,itches skintexturewasimproved. wereremoved. Q98 female 200s chronic C onceaday immediatelyafterapplication,erythema eczema andscaleswereremitted. Q99 male 20s atopic C onceaday After2minutes,rednesswasimproved. After1minute,iches dermatitis After3minutes,erythema,infiltration wereeliminated. andpapulewereremitted.After1 After1application, application,theeffectlastedfor6 noitchesfor6days dayswithnorecurrence. thereafter. Q100 female 50s prurigo C onceaday After3minutes,nodeswereflatened, Icheswereeliminated nodularis, scratchwoundbecamedryandepithelized immediatelyafter eczema withnomoreexudate. application. 8 SEQIDNO39: Q101 female 50s prurigo C onceaday After3minutes,nodeswereflattened. (CFSRKMDRISSSSGLGC) nodularis, eczema Q102 female 10s chroniceczema C onceaday After2minutes,anintractableeczema thatcannotberemittedbyapplyingstrong steriodointmentswasremitted,and erythema,scalesandpapulewerereduced. Q103 female 50s eczema C onceaday After2minutes,erythema,scalesand pigmentationwereremitted.Skingotfull andsoft,withfreshcolor. Q104 female 20s atopic D onceaday ScratchwoundwasepithelizedExudatewas Icheswereeliminated dermatitis driedadlichenificationwasremitted. immediatelyafter erythemaandinfiltrationwereremitted. application. Q105 male 20s atopic B onceaday After3minutes,erythemaandinfiltration After2minutes, dermatitis wereremitted.With1application,the itchesandtingling effectpersistedfor1weekwithno painwereremoved. recurrence. Theeffectpersisted for7dayswithno itches. SEQIDNO40: Q106 male 40s atopic C onceaday After2minutes,erythema,infiltration Icheswereeliminated (CFTRKMDRISSSSGLGC) dermatitis andlichenificationremitted. immediatelyafter application. Q107 female 20s roughened C 1application After1minute,dryskinwasimproved, lips crackswerelessdeepandapithelized. Q108 female 40s roughened B onceaday After3minutes,rednessandroughness skin wereimproved. Q109 female 30s healthyskin C onceaday After3minutes,dark-reddishskintone wasimprovedtoturnfreshcolor. Dryness,tautnessmrawnessandhotflash ofskinwereremoved.With1application theeffectpersistedfor1week. 9 SEQIDNO41: Q110 male 10s atopic C onceaday immediatelyafterapplication,erythema Icheswereeliminated (CFGRKMDRISSTSGLGC) dermatitis/ andinfiltrationofeczemawereremitted. immediatelyafter application. acne Theeffectswereexhibitedremarkably fast.After3minutes,erythemawas furtherremitted.Acnewasdriedand reducedinsize. Q111 male 20s atopic C onceaday Immediatelyafterapplication,erythema Icheswereeliminated dermatitis andinfiltrationwereremitted.After immediatelyafter 20minutes,deepscratchwoundwas application. epithelized. Q112 female 40s roughened B 1application After1minute,skintautnessandrawness lips wereimprovedandskingotmoisturized. Q113 female 50s atopic B onceaday Ichesstoppedin30seconds.Redness dermatitis andedemawereremittedin1minute. Q114 female 10s roughened B onceaday After1minute,rednesswasremitted. After3minutes,skin skin/acne After3minutes,skinroughness,zits, tautnesswasremoved. roughandsoftskintexturewereimproved, skingotfullenditstexturewasfinely improved.Acnebecamedryandreducedin size. Q115 female 40s roughened B onceaday Rawnessvanishedin1minute.Skinwas skin moisturized.After2minutes,redness, roughnessandstiffnesswereremarkably improved.Byusingfor1weel,skin becamebrighter,flabbinesswasimproved. SEQIDNO42: Q116 female 40s acne C onceaday After1minute,rednesswasremitted. (CFGRKMDRISSASGLGC) Q117 male 10s atopic B onceaday After3minutes,erythemaand Icheswereeliminated dermatitis infiltrationwereremitted. immediatelyafter application. Q118 female 50s eczema C onceaday After3minutes,erythemaanddryskin remitted. SEQIDNO43: Q119 male 40s atopic C onceaday After2minutes,nodiscomfort,felt After3minutes, (CFGRKMDRISSSTGLGC) dermatitis fresh.Erythema,infiltrationand roughnessand lichenificationwereremitted. stiffnesswere removed,anditches vanished. Q120 female 30s roughened C onceaday 1minuteafterapplication,dryskin lips wasimprovedcracksbecamelessdeep andepithelized. Q121 female 40s roughened B onceaday After3minutes,rednessanddryskin skin wereimproved.Skingotmoisturized promptly,andskintexturewasimproved. Skinrawness,stiffnessandirritation wereeliminated.Poresbecameless distinguised. Q122 female 40s rosacae/acne C onceaday After7minutes,acnewasreduceand becamedry. Q123 female 30s eczema C onceaday After1minute,rednessandinfiltration Icheswereeliminated wereremitted. immediatelyafter application. 10 SEQIDNO44: Q124 female 60s healthyskin B onceaday Skingotmoisturizedpromptly,wrinkles (CFGRKMDRISSSAGLGC) becamelessdeep,skintexturewasimproved. Byusingfor4weeks,largewrinklesand nasolabialfoldbecamelessdeep.Skin resilenceandflabinesswereimproved. Spotsfaded.Skintonebecamebrighter. Q125 female 80s eczema C onceaday After1minute,erythemaandinfiltration Icheswereremitted wereremitted. immediatelyafter application. SEQIDNO45: Q126 female 40s roughened B onceaday After2minutes,dryness,tautnessand (CFSRRMDRISSSSGLGC) skin rawnessofskinwereeliminated,skingot full,andwrinklesbecamelessdeep.Skin mainedelasticity.Byusingfor5days, flabbinesswasimproved.Skinbecame brighter,nasolabialfoldbecameless deep.Byusingfor4weeks,nasolabial foldbecamelessdeep.Skinresilence andflabbinesswereimproved.Spotsfaded andskintonebecamebrighter. SEQIDNO46: Q127 female 40s eczema C onceaday After2minutes,erythema,scalesand (CFGRKMDRISSTSGLGC) infiltrationwereremitted.Skingot softened. SEQIDNO47: Q128 female 30s chroniceczema C onceaday Immediatelyafterapplication,erythema (CFGRKMDRISSSTGLGC) andscaleswereremitted. SEQIDNO48: Q129 male 50s eczema C onceaday After2minutes,erythema,scalesand (CFGRKMDRISSSSGIGC) infiltrationwereremitted.Skingot moisturized. SEQIDNO49: Q130 male 30s chroniceczema C onceaday Immediatelyafterapplication,erythema (CFGRKMDRISSSSGLAC) andscaleswereremitted. SEQIDNO50: Q131 male 50s eczema C onceaday After2minutes,erythema,scalesand (CFGRRMDRISSSTGLGC) infiltrationwereremitted. SEQIDNO51: Q132 female 60s eczema B onceaday After3minutes,erythemaand (CFGRRMDRISSSTGLGC) infiltrationwereremitted. SEQIDNO52: Q133 female 50s eczema C onceaday After2minutes,erythema,scalesand (CFGRRMDRISSSSGIGC) infiltrationwereremitted.Skingot moisturized. SEQIDNO53: Q134 female 20s eczema C onceaday After1minute,erythemaandinfiltration (CFGRRMDRISSSSGLAC) wereremitted. SEQIDNO54: Q135 female 30s roughened B onceaday After2minutes,dryness,tautnessand (CFSRKMDRISSTSGIGC) skin rawnessofskinwereeliminated,skin gotfullandwrinklesbecamelessdeep. Skingainedelasticity. SEQIDNO54: Q136 female 50s eczema C onceaday After2minutes,erythema,scaleand (CFSRKMDRISSTSGIGC) infiltrationwereremitted. 11 SEQIDNO56: Q137 male 20s roughened B onceaday After2minutes,dryness,tautnessand (CFGRKMDRISSSTGIGC) skin rawnessofskinwereelimnated,skingot full,andwrinklesbecamelessdeep. Skingainedelasticity. SEQIDNO57: Q138 female 40s eczema C onceaday After2minutes,erythema,scalesand (CFGRKMDRISSSTGLAC) infiltrationwereremitted,scratch woundwasepithelizedanddried. SEQIDNO58: Q139 female 30s eczema C onceaday After2minutes,erythema,scales, (CFGRKMDRISSSSGIAC) papuleandinfiltrationwereremitted. SEQIDNO59: Q140 male 60s roughened B onceaday After2minutes,dryness,tautnessand (CFGRKMDRISSTSGLGC) skin rawnessofskinwereeliminated.Skin gainedelasticity,wrinklesbecameless deep. SEQIDNO60: Q141 female 50s chronic C onceaday Immediatelyafterapplication,erythema (CFGRRMDRISSSTGLGC) eczema andscaleswereremitted. SEQIDNO61: Q142 female 30s chronic C onceaday Immediatelyafterapplication,erythema (CFSRRMDRISSSSGIGC) andscaleswereremitted. SEQIDNO62: Q143 female 40s roughened B onceaday After2minutes,dryness,tautnessand (CFSRRMDRISSSSGLAC) skin rawnessofskinwereeliminated,skin gotfull,andwrinklesbecamelessdeep. Skingainedelasticity. SEQIDNO63: Q144 male 50s eczema B onceaday After3minutes,erythemaandscales (CFSRKMDRISSTSGIGC) infiltrationwereremitted,andskingot softened. SEQIDNO64: Q145 male 40s chronic C onceaday Immediatelyafterapplication,erythema (CFSRKMDRISSTSGIGC) eczema andscaleswereremitted. SEQIDNO65: Q146 female 40s roughened C onceaday After1minute,skingotmoisturizedand (CFGRKMDRISSSTGLAC) skin resilent,skintexturewasimproved. SEQIDNO66: Q147 male 50s eczema C onceaday After2minutes,erythema,scalesand (CFGRKMDRISSSSGIAC) infiltrationwereremitted. SEQIDNO67: Q148 female 70s roughened B onceaday After2minutes,dryness,tautnessand (CFSRRMDRISSTSGIGC) skin rawnessofskinwereeliminated,skin gotfull,andwrinklesbecamelessdeep. Skingainedelasticity. SEQIDNO68: Q149 female 30s chronic C onceaday Immediatelyafterapplication,erythema (CFGRRMDRISSTSGLAC) eczema andscaleswereremitted. SEQIDNO69: Q150 female 20s eczema C onceaday After2minutes,erythema,scales, (CFSRRMDRISSTSGIGC) infiltrationwereimproved. 12 SEQIDNO70: 151 female 60s chronic C onceaday immediatelyafterapplication,erythema (CFGRRMDRISSSTGLAC) eczema andscaleswereremitted. 152 female 30s roughened C onceaday Rednessvanishedin30seconds.After1 skin minute,skintextureimprovedfinely. SEQIDNO71: Poresbecamelessdistinguished. (CFSRRMDRISSTSGIGC) 153 male 50s roughened A onceaday After2minutes,dryness,tautnessand skin rawnessofskinwereeliminated,skin gotfullandwrinklesbecamelessdeep. Skingainedelasticity. SEQIDNO72: 154 male 50s chronic C onceaday Immediatelyafterapplication,erythema, (CFSRRMDRISSTSGLAC) eczema scalesandpapulewereremitted. SEQIDNO73: 155 male 20s chronic C onceaday Immdediatelyafterapplication,erythema (CFSRRMDRISSSTGIGC) eczema andscaleswereremitted. SEQIDNO74: 156 female 30s roughened C onceaday After1minute,rawnessandhotflash (CFGRRMDRISSSTGLAC) skin wereeliminated,skingotmoisturized anditstextureimproved,wrinklesfaded andskingainedelasticity. 157 male 60s chronic D onceaday Icheswereeliminatedimmediatelyafter eczema application.Rednesswasremitted. SEQIDNO75: 158 female 60s eczema C onceaday After2minutes,erythema,scalesm (CFGRRMDRISSSSGIAC) papule,infiltrationwereremitted. SEQIDNO43: 258 male 40s atopic C onceaday After2minutes,crackswereepithelized (CFGRKMDRISSSTGLGC) dermatitis withnomorepainsanditches.Erythema and andscaleswerealsoremitted. SEQIDNO19: (CFGRRMDRISSSSLGC) SEQIDNO43: 358 male 40s eczema onceaday After2minutes,woundwasepithelized (CFGRKMDRISSSTGLGC) quickly,anderythema,scalesand and infiltrationwereremarkablyremitted. SEQIDNO99: In SEQ ID NOS 16-75, 1st Cys and 17th Cys from left form a disulfide bond.

[0293] The result described above demonstrated that the cyclic peptide of the invention exhibits a similar effect as the cyclic peptide expressed by Formula I-a even if some amino acids of the cyclic peptide have been replaced. Moreover, a similar effect could be obtained by applying the replaced cyclic peptide in a mixture as long as an effective amount of the cyclic peptide is applied.

[0294] Specifically, in subjects having atopic dermatitis or eczema, effects were observed immediately after applying 3 g/ml to 500 g/ml to affected site, including elimination of itches, epithelialization at infiltrated site, papule or scratch, termination of exudation, epithelialization of a crack. Moreover, a single application induced a persistent effect for a week or more.

[0295] When it was applied to healthy or roughened skin, it exerted a moisturizing effect immediately or within a few minutes after application, improving dryness, providing and keeping skin and mucosa with moderate elasticity and flexibility, softening skin, giving skin or mucosa resilience and firmness. Also, fine wrinkles and flabbiness are improved at the applied site, and dullness and spots are further faded. A single application induced a persistent effect for a week or more.

9.3 Confirming Nasal Effect

[0296] The obtained cyclic peptide was prepared in physiological saline at 3, 15, 30, 100 or 500 g/ml, and 0.1 ml each was applied to nasal cavity. Each of formulations A to E in the table indicates the concentration of the cyclic peptide at 3, 15, 30, 100 or 500 g/ml, respectively. The absence of the effect upon the application of physiological saline without the peptide confirmed the effect was not a placebo effect.

TABLE-US-00031 TABLE13-1 SEQIDNO (aminoacid sequence)of Diagnostic Formu- Diagnosticimpressionon Itches cyclicpeptides ID Sex Age impression lation Treatment skinafterapplication afterapplication SEQIDNO16: P1 female 10's allergic C onceaday,0.1ml Fromimmediatelyafternasal Itcheswereremitted (CFVRKMDRISSS rhinitis appliedtoeach application,andnosegot immediatelyafternasal SGLGC) nasalcavity cleared,rhinorrheastopped. application,andeliminated Therewasnoirritationg after1minute. symptoms,andtheeffect persistedforalldayby1 nasalapplication. SEQIDNO17: P2 male 30's allergic D onceaday,0.1ml Fromimmediatelyafternasal Itcheswereremitted (CFGQKMDRISSS rhinitis appliedtoeach application,andnosegot immediatelyafternasal SGLGC) nasalcavity cleared,rhinorrheastopped. application,andeliminated Therewasnoirritationg after1minute. symptoms,andtheeffect persistedforalldayby1 nasalapplication. SEQIDNO18: P3 male 50's allergic C onceaday,0.1ml After3minutes,andnose (CFGHKMDRISSS rhinitis appliedtoeach gotcleared,rhinorrhea SGLGC) nasalcavity stopped.Therewasno irritationgsymptoms,and theeffectpersistedforall dayby1nasalapplication. SEQIDNO19: P4 female 40's chronic B onceaday,0.1ml After1minute,andnosegot (CFGRRMDRISSS rhinitis, appliedtoeach cleared,nasaldischarge SGLGC) perennial nasalcavity stopped.Therewasno rhinitis irritatingsymptoms. P5 female 70's allergic B onceaday,0.1ml After2minutes,andnose Itcheswereremitted rhinitis appliedtoeach gotcleared,andnonasal immediatelyafternasal nasalcavity dischargecameoutby application. blowingnose. SEQIDNO20: P6 female 40's allergic B onceaday,0.1ml After1minute,nasal (CFGRKLDRISSS rhinitis appliedtoeach obstructionwasimproved, SGLGC) nasalcavity andnosegotcleared.The effectpersistedforall day. SEQIDNO21: P7 female 20's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKIDRISSS rhinitis appliedtoeach cleared.Nasaldischarge SGLGC) nasalcavity stopped. SEQIDNO22: P8 female 50's allergic E onceaday,0.1ml Immediatelyafternasal Immediatelyafternasal (CFGRKMDRVSSS rhinitis appliedtoeach application,andnosegot application,restlessitches SGLGC) nasalcavity cleared. andsneezingstopped. P9 female 40's chronic C onceaday,0.1ml After20seconds,andnose rhinitis, appliedtoeach gotcleared.Nonasal perennial nasalcavity dischargeafter1minute. rhinitis Therewasnoirritating symptoms. SEQIDNO23: P10 female 30's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRIGSS rhinitis appliedtoeach cleared.Nasaldischarge SGLGC) nasalcavity stopped. SEQIDNO24: P11 female 20's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRISAS rhinitis appliedtoeach cleared.Nasaldischarge SGLGC) nasalcavity stopped. SEQIDNO25: P12 female 20's allergic A onceaday,0.1ml After2.5minutes,andnose (CFGRKMDRISSQ rhinitis appliedtoeach gotcleared. SGLGC) nasalcavity SEQIDNO26: P13 female 50's allergic C onceaday,0.1ml After1minute,nosegot (CFGRKMDRISSV rhinitis appliedtoeach clearedandrefreshed.After SGLGC) nasalcavity 2minutes,nonasal dischargebyblowing. SEQIDNO27: P14 male 20's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRISSL rhinitis appliedtoeach cleared. SGLGC) nasal SEQIDNO28: P15 female 40's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRISSI rhinitis appliedtoeach cleared.Nasaldischarge SGLGC) nasalcavity stopped. SEQIDNO29: P16 male 50's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRISSM rhinitis appliedtoeach cleared.Nasaldischarge SGLGC) nasalcavity stopped. SEQIDNO30: P17 female 30's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFGRKMDRISSS rhinitis appliedtoeach stopped. VGLGC) nasalcavity SEQIDNO31: P18 female 10's allergic D onceaday,0.1ml After30seconds,rhinorrhea (CFGRKMDRISSS rhinitis appliedtoeach stopped.After1minute,no SRLGC) nasalcavity nasaldischargebyblowing. SEQIDNO32: P19 male 40's allergic C onceaday,0.1ml After1minute,nasal (CFGRKMDRISSS rhinitis appliedtoeach obstructionwasimproved, SGMGC) nasalcavity andnosegotcleared.The effectpersistedforall day. P20 female 40's chronic C onceaday,0.1ml Immediatelyafternasal rhinitis, appliedtoeach applicationnosegot perennial nasalcavity cleared.Theeffectwas rhinitis remarkableandpersisted for7daysafter1nasal application. SEQIDNO33: P21 female 40's allergic C onceaday,0.1ml After30seconds,rhinorrhea Sneezinganditchesindeep (CFGRKMDRISSS rhinitis appliedtoeach stopped.After1minute,no insidenosestopped. SGIGC) nasalcavity nasaldischargebyblowing. SEQIDNO34: P22 female 40's allergic C onceaday,0.1ml After2.5minutes,andnose Noirritatingsymptoms. (CFGRKMDRISSS rhinitis appliedtoeach gotcleared. SGVGC) nasal SEQIDNO35: P23 female 30's allergic C onceaday,0.1ml After1minute,nasal (GFGRKMDRISSS rhinitis appliedtoeach obstructionwasimproved, SGAGC) nasalcavity andnosegotcleared.The effectpersistedforall day. P24 female 40's chronic D onceaday,0.1ml Immediatelyafternasal rhinitis, appliedtoeach application,nosegot perennial nasalcavity refreshedandcleared. rhinitis SEQIDNO36: P25 female 40's allergic C onceaday,0.1ml Immediatelyafternasal Immediatelyafternasal (CFGRKMDRISSS rhinitis appliedtoeach application,andnosegot application,itcheswere SGLSC) nasalcavity cleared. removed.Noirritating symptoms. SEQIDNO37: P26 female 40's allergic C onceaday,0.1ml After30seconds,rhinorrhea Immediateeffecttoremove (CFGRKMDRISSS rhinitis appliedtoeach stopped.Nosegotcleared itches. SGLAC) nasalcavity andfeltgood.Othernose dropscausesthissubjectto sneezeduetosomeirritant, buttheinventivenosedrop hadnoirritation.The effectpersistedfor1week by1nasalapplication. P27 female 40's chronic C onceaday,0.1ml Nosegotclearedand rhinitis, appliedtoeach rhinorrheastoppedin30 perennial nasalcavity seconds. rhinitis SEQIDNO38: P28 male 30's chronic C onceaday,0.1ml After2minutes,rhinorrhea (CFARKMDRISSS rhinitis appliedtoeach stopped.Nasalobstruction SGLGC) nasalcavity wasimprovedtoallownasal breathing. SEQIDNO39: P29 male 40's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRKMDRISSS rhinitis appliedtoeach application,nosegot SGLGC) nasalcavity refreshedandcleared.No nasaldischargeafter30 seconds. SEQIDNO40: P30 female 30's allergic C onceaday,0.1ml After2.5minutes,andnose (CFTRKMDRISSS rhinitis appliedtoeach gotcleared. SGLGC) nasal SEQIDNO41: P31 female 20's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFGRKMDRISST rhinitis appliedtoeach stopped.Therewasno SGLGC) nasalcavity irritatingsymptoms. SEQIDNO42: P32 female 10's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRKMDRISSA rhinitis appliedtoeach application,andnosegot SGLGC) nasalcavity cleared. SEQIDNO43: P33 male 20's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRKMDRISSS rhinitis appliedtoeach application,andnosegot TGLGC) nasalcavity cleared.After1minute,no nasaldischargeevenby twistingnose. SEQIDNO44: P34 male 40's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFGRKMDRISSS rhinitis appliedtoeach stopped.Therewasno AGLGC) nasalcavity irritatingsymptoms. SEQIDNO45: P35 male 60's allergic C onceaday,0.1ml After1minute,andnosegot (CFSRRMDRISSS rhinitis appliedtoeach cleared. SGLGC) nasal SEQIDNO47: P36 male 50's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFSRKMDRISSS rhinitis appliedtoeach stopped.Therewasno TGLGC) nasalcavity irritatingsymptoms. SEQIDNO48: P37 female 50's allergic C onceaday,0.1ml After1minute,andnosegot (CFSRKMDRISSS rhinitis appliedtoeach cleared.Nasaldischarge SGIGC) nasalcavity stopped. SEQIDNO49: P38 female 40's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRKMDRISSS rhinitis appliedtoeach application,andnosegot SGLAC) nasalcavity cleared. SEQIDNO50: P39 female 10's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRRMDRISST rhinitis appliedtoeach application,andnosegot SGLGC) nasalcavity cleared. SEQIDNO51: P40 male 20's chronic C onceaday,0.1ml After1.5minutes,nasal (CFGRRMDRISSS rhinitis appliedtoeach obstructionwasimproved, TGLGC) nasalcavity andnosegotcleared. SEQIDNO52: P41 male 30's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFGRRMDRISSS rhinitis appliedtoeach stopped.Therewasno SGIGC) nasalcavity irritatingsymptoms. SEQIDNO53: P42 male 40's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRRMDRISSS rhinitis appliedtoeach application,andnosegot SGLAC) nasalcavity cleared. SEQIDNO54: P43 female 30's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRKMDRISST rhinitis appliedtoeach application,andnosegot SGIGC) nasalcavity cleared. SEQIDNO55: P44 male 50's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRISST rhinitis appliedtoeach cleared.Nasaldischarge SGLAC) nasalcavity stopped. SEQIDNO56: P45 male 30's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRKMDRISSS rhinitis appliedtoeach application,andnosegot TGIGC) nasalcavity cleared. SEQIDNO57: P46 female 40's allergic C onceaday,0.1ml After1minute,andnosegot (CFGRKMDRISSS rhinitis appliedtoeach cleared.Nasaldischarge TGLAC) nasalcavity stopped. SEQIDNO58: P47 female 40's allergic C onceaday,0.1ml After30seconds,rhinorrhea Itcheswereremovedin (CFGRKMDRISSS rhinitis appliedtoeach stopped,withnomore 30seconds. SGIAC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO59: P48 female 20's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRRMDRISST rhinitis appliedtoeach application,andnosegot SGLGC) nasalcavity cleared. SEQIDNO60: P49 female 40's allergic C onceaday,0.1ml After1minute,andnosegot (CFSRRMDRISSS rhinitis appliedtoeach cleared.Nasaldischarge TGLGC) nasalcavity stopped. SEQIDNO61: P50 male 30's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRRMDRISSS rhinitis appliedtoeach application,andnosegot SGIGC) nasalcavity cleared. SEQIDNO62: P51 female 40's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFSRRMDRISSS rhinitis appliedtoeach stopped,withnomore SGLAC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO63: P52 female 30's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRKMDRISST rhinitis appliedtoeach application,andnosegot SGIGC) nasalcavity cleared. SEQIDNO64: P53 male 60's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFSRKMDRISSS rhinitis appliedtoeach stopped,withnomore TGIGC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO65: P54 female 30's allergic C onceaday,0.1ml After1minute,andnosegot (CFSRKMDRISSS rhinitis appliedtoeach cleared.Nasaldischarge TGLAC) nasalcavity stopped. SEQIDNO66: P55 female 30's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRKMDRISSS rhinitis appliedtoeach application,andnosegot SGIAC) nasalcavity cleared. SEQIDNO67: P56 female 40's allergic C onceaday,0.1ml After1minute,andnose (CFGRRMDRISST rhinitis appliedtoeach gotcleared.Nasal SGIGC) nasalcavity dischargestopped. SEQIDNO68: P57 male 60's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFGRRMDRISST rhinitis appliedtoeach stopped,withnomore SGLAC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO69: P58 male 20's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRRMDRISSS rhinitis appliedtoeach application,andnosegot TGIGC) nasalcavity cleared. SEQIDNO70: P59 female 40's allergic C onceaday,0.1ml Immediatelyafternasal (CFGRRMDRISSS rhinitis appliedtoeach application,andnosegot TGLAC) nasalcavity cleared. SEQIDNO71: P60 female 30's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFSRRMDRISST rhinitis appliedtoeach stopped,withnomore SGIGC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO72: P61 female 40's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFSRRMDRISST rhinitis appliedtoeach stopped,withnomore SGLAC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO73: P62 female 20's allergic C onceaday,0.1ml Immediatelyafternasal (CFSRRMDRISSS rhinitis appliedtoeach application,andnosegot TGIGC) nasalcavity cleared. SEQIDNO74: P63 female 40's allergic C onceaday,0.1ml After1minute,rhinorrhea (CFSRRMDRISSS rhinitis appliedtoeach stopped,withnomore TGLAC) nasalcavity sneezing.Therewasno irritatingsymptoms. SEQIDNO75: P64 male 20's allergic C onceaday,0.1ml After1minute,andnose (CFSRRMDRISSS rhinitis appliedtoeach gotcleared.Nasal SGIAC) nasalcavity dischargestopped. In SEQ ID NOS 16-75, 1st Cys and 17th Cys from left form a disulfide bond.

[0297] The result described above demonstrated that the cyclic peptide of the invention exhibits a similar effect as the cyclic peptide expressed by Formula I-a even if some amino acids of the cyclic peptide have been replaced.

[0298] Specifically, when it was applied to nasal cavity mucosa and/or paranasal cavity mucosa, it can improve or prevent various symptoms associated with rhinitis such as nasal obstruction, rhinorrhea, sneezing and pruritus. Rhinitis also includes, without being particularly limited, for example, infectious rhinitis including acute rhinitis and chronic rhinitis; hypersensitive non-infectious rhinitis including combined (flower hypersensitivity) rhinitis, rhinitis with rhinorrhea, congestive rhinitis, edematous rhinitis and dry-nose type rhinitis; irritant-induced rhinitis including physical rhinitis, chemical rhinitis and radiation rhinitis; and atrophic rhinitis and idiopathic granulomatous rhinitis; sinusitis such as acute sinusitis, chronic sinusitis (maxillary empyema), eosinophilic sinusitis and paranasal cavity mycosis. The external preparation can be used for the purpose of treating or preventing one ore more of the above.

[0299] Combined rhinitis includes, for example, allergic rhinitis including perennial allergic rhinitis and seasonal allergic rhinitis, and nonallergic rhinitis including vasomotor (essential) rhinitis and eosinophilic rhinitis. Rhinitis with rhinorrhea includes, for example, gustatory rhinitis, cold air-induced rhinitis and senile rhinitis.

[0300] Congestive rhinitis includes, for example, drug-induced rhinitis, psychogenic rhinitis, gestational rhinitis, endocrine rhinitis and cold rhinitis.

[0301] Edematous rhinitis includes, for example, aspirin-hypersensitive rhinitis.

[0302] Among those mentioned above, it exerts an excellent effect specifically on hypersensitive non-infectious rhinitis, irritant-induced rhinitis and sinusitis, preferably on hypersensitive non-infectious rhinitis and chronic sinusitis, more preferably on combined (flower hypersensitivity) rhinitis, rhinitis with rhinorrhea, congestive rhinitis, edematous rhinitis and dry-nose type rhinitis, and chronic sinusitis. Therefore, the external preparation of the present invention can be used for the purpose of treating or preventing at least one or more rhinitis selected from the above-mentioned group.

[0303] In terms of its symptoms, for its effects as described above, it can be used for any of rhinitis with sneezing/rhinorrhea, rhinitis with nasal obstruction and rhinitis with both symptoms.

[0304] Immediately after a single application of 0.1 ml of 3 g/ml to 500 g/ml preparation was applied to the affected site in each nasal cavity, nasal discharge was stopped, nasal obstruction was improved, itches in nasal cavity mucosa or sneezing were stopped (immediate effect). A single application of a nasal drop induced persistent effects lasting for one day, or for a week or more in some cases (long-lasting effect). In addition, because it causes no local irritation at all, it can be used safely even in a subject who is not desired to use a nasal drop comprising rhinitis therapeutic that has conventionally been used such as steroid drug due to its strong local irritation.

[0305] Examples of multiple replaceable amino acids in the cyclic peptide of the invention are summarized below, based on working examples as described above, without not being limited thereto.

TABLE-US-00032 TABLE32 CandidatemultiplereplacementsofBring Number Exampleof of Resi- Resi- Resi- Resi- replaced SEQ replace- due Amino due Amino due Amino due Amino aminoacid ID ment ID acid ID acid ID acid ID acid sequence NO: 2 3 A,S 5 R CFSRRMDRISSSSGLGC 45 3 A,S 12 Q,T CFSRKMDRISSTSGLGC 46 3 A,S 13 T CFSRKMDRISSSTGLGC 47 3 A,S 15 M,I, CFSRKMDRISSSSGIGC 48 V,A 3 A,S 16 A,S CFSRKMDRISSSSGLAC 49 5 R 12 Q,T CFGRRMDRISSTSGLGC 50 5 R 13 T CFGRRMDRISSSTGLGC 51 5 R 15 M,I, CFGRRMDRISSSSGIGC 52 V,A 5 R 16 A,S CFCRRMDRISSSSGLAC 53 12 Q,T 15 M,I, CFGRKMDRISSTSGIGC 54 V,A 12 Q,T 16 A,S CFGRKMDRISSTSGLAC 55 13 T 15 M,I, CFGRKMDRISSSTGIGC 56 V,A 13 T 16 A,S CFGRKMDRISSSTGLAC 57 15 M,I, 16 A,S CFGRKMDRISSSSGIAC 58 V,A 3 3 A,S 5 R 12 Q,T CFSRRMDRISSTSGLGC 59 3 A,S 5 R 13 T CFSRRMDRISSSTGLGC 60 3 A,S 5 R 15 M,I,V,A CFSRRMDRISSSSGIGC 61 3 A,S 5 R 16 A,S CFSRRMDRISSSSGLAC 62 3 A,S 12 Q,T 15 M,I,V,A CFSRKMDRISSTSG1GC 63 3 A,S 13 T 15 M,I,V,A CFSRKMDRISSSTGIGC 64 3 A,S 13 T 16 A,S CFSRKMDRISSSTGLAC 65 3 A,S 15 M,I, 16 A,S CFSRKMDRISSSSGLAC 66 V,A 5 R 12 Q,T 15 M,I,V,A CFGRRMDRISSTSGIGC 67 5 R 12 Q,T 16 A,S CFGRRMDRISSTSGIAC 68 5 R 13 T 15 M,I,V,A CFGRRMDRISSSTGIGC 69 5 R 13 T 16 A,S CFGRRMDRISSSTGLAC 70 5 R 15 M,I, 16 A,S CFGRRMDRISSSSGLAC 71 V,A 4 3 A,S 5 R 12 Q,T 15 M,I, CFSRRMDR1SSTSGIGC 72 V,A 3 A,S 5 R 12 Q,T 16 A,S CFSRRMDRISSTSGLAC 73 3 A,S 5 R 13 T 15 M,I, CFSRRMDRISSSTGIGC 74 V,A 3 A,S 5 R 13 T 16 A,S CFSRRMDRISSSTGLAC 75 3 A,S 5 R 15 M,I,V,A 16 A,S CFSRRMDRISSSSGIAC 76