JP5654719B2 - Synergistic combination therapy to treat vitiligo - Google Patents

Description

  The present invention relates to a method of treating vitiligo using a synergistic formulation.

  Vitiligo / leucoderma is an acquired depigmentation of the skin characterized by patchy loss of pigment that becomes progressive over time. About 1% of the world population suffers from this disease. Traditional treatments for vitiligo mainly include topical / oral psoralen followed by exposure to ultraviolet A radiation (PUV-A) or photochemotherapy with topical / oral steroids. PUV-A therapy is probably the mainstay in the treatment of vitiligo. However, re-pigmentation is obtained only in about 50% of cases. More in the patients who respond to PUV-A, more vitiligo can be re-pigmented, albeit only in part, and the remaining vitiligo can be treated with PUV-A therapy even after prolonged treatment. It remains unresponsive. The repigmentation in the above treatment is the result of an increase in the cells, melanocytes, that produce pigment melanin in the skin. An increase in melanocytes in response to the treatment occurs either from the edge of the vitiligo or from migration / diffusion to the pigmented hair follicle and its vitiligo.

  Basic fibroblast growth factor (bFGF), also known as FGF2, because it contains a number of basic amino acid residues (lysine, arginine and histidine), is a potent against various cell types including melanocytes. A mitogen. Both human and bovine bFGF were isolated and the gene expressing this product was sequenced and cloned. In addition, bFGF has been found to be expressed in a wide variety of tissue types, including placenta, keratinocytes and fibroblasts. bFGF or its agonist peptides have been tested in various phases of clinical trials in human volunteers in India and repigmented approximately 80% of volunteers with stable generalized and segmental vitiligo It was found to be successful. Applicable patents describing bFGF or agonist peptides and formulations for penetrating them into complete skin include US Pat. No. 6,143,723, Australian Patent No. 722626, Indian Patent Nos. 185613, 186437 and 185703. Is included.

  Vitiligo is a pigmented disease associated with patchy loss of the skin pigment melanin (Ramaiah. A, Puri. N, Mojamdar M, A new hypothesis for the ethology of vitiligo, “A new hypothesis on the etiology of vitiligo”, Acter Dol. Stockholm), 1989, 69, 323-327), the deprivation or reduced level of basic fibroblast growth factor (bFGF) -like mitogen for melanocytes in the skin of vitiligo patients is unknown It is hypothesized that this would cause the loss of melanocytes in the skin producing vitiligo. Basic fibroblast growth factor (bFGF), also known as FGF2, is a potent mitogen for a variety of cell types including melanocytes. Both human and bovine bFGF have been isolated and the gene expressing this product has been sequenced and cloned. In addition, bFGF has been found to be expressed in a wide variety of tissue types including pituitary, brain and adrenal corpus luteum, retina, kidney, placenta as well as keratinocytes, fibroblasts. The above assumption that bFGF-like mitogens are reduced for their levels in vitiligo resulting in loss of melanocytes and pigment melanin in vitiligo skin has recently been studied by others (Moretti S et al, Insight into the pathogenesis of vitiligo, (Imbalance of epidemitical cytokines at sites of relations, Pig. Cell. Res 2002, 15).

According to the present invention, a method for synergistic combinatorial therapy for the treatment of generalized and segmental vitiligo, with psoralen and UV-A (PUV-A therapy) / PUV-ASOL There is provided a method comprising topical application of an effective amount of a composition comprising 0.02-5% w / w of at least one peptide selected from the group consisting of SEQ ID NOS: 1-8 described herein. .
According to an embodiment, a method for synergistic combination therapy for the treatment of generalized and segmental vitiligo comprising:
0.02-5% w / w of bFGF amino acids 106-115 (SEQ ID NO: 1), bFGF amino acids 106-120 (SEQ ID NO: 5), bFGF amino acids 1-24 (SEQ ID NO: 6) and SEQ ID NOs: 2, 3, At least one peptide selected from the group consisting of 4, 7 and 8;
10-50% w / w solvent,
Topically applying an effective amount of a composition comprising 10-40% w / w glycol, and 10-40% w / w of at least one penetration enhancer ,
The composition is administered with psoralen and UV-A (PUV-A therapy) / PUVASOL,
A method is provided.

A method for a synergistic combination therapy for the treatment of vitiligo that does not respond to PUV-A therapy, comprising continuing oral ingestion of psoralen and exposure to UV-A from SEQ ID NOs: 1-8 A method comprising topical application of an effective amount of a composition comprising 0.02-5% w / w of at least one peptide selected from the group consisting of:
According to an embodiment, a method for a synergistic combination therapy for the treatment of vitiligo that does not respond to PUV-A therapy, comprising:
0.02-5% w / w of bFGF amino acids 106-115 (SEQ ID NO: 1), bFGF amino acids 106-120 (SEQ ID NO: 5), bFGF amino acids 1-24 (SEQ ID NO: 6) and SEQ ID NOs: 2, 3, At least one peptide selected from the group consisting of 4, 7 and 8;
10-50% w / w solvent,
Topically applying an effective amount of a composition comprising 10-40% w / w glycol, and 10-40% w / w of at least one penetration enhancer ,
The composition is administered with continued ingestion of psoralen and exposure to UV-A;
A method is provided.

A method for synergistic combination therapy for the treatment of fast spreading vitiligo, wherein 0.02 of at least one peptide selected from the group consisting of SEQ ID NOs: 1-8 in the above formulation A method comprising topical application of an effective amount of a composition comprising ˜5% w / w. The above composition is administered with steroid therapy.

A method for synergistic combination therapy for the treatment of vitiligo, have accompanied the surgical procedure, in the above formulation, of at least one peptide selected from the group consisting of SEQ ID NO: 1-8 0.02 A method comprising topical application of an effective amount of a composition comprising ˜5% w / w.

A method for synergistic combination therapy for the treatment of vitiligo comprising 10-50% w / w solvent, 10-40% w / w glycol and 10-40% w / w penetration A method comprising topical application of an effective amount of a composition further comprising a penetration enhancer.

Topical application of the peptide in the above formulation is effective in 80% or more cases of stable generalized or segmental vitiligo. Even better results with a repigmentation rate and success rate of 80% or more appear to be achieved when combined with traditional therapies such as PUV-A therapy for vitiligo. In addition, bFGF peptide lotion therapy is not effective in preventing the rapid spread of vitiligo, and perhaps synergistic combination therapy is traditionally used in the case of fast diffuse vitiligo cases It appears when used in combination with steroid therapy. Similarly, the topical application of the peptide lotion therapy is advantageously used with any other therapy including surgery or other therapy related to immunomodulators.

A synergistic combination therapy for the treatment of fast-diffusing vitiligo cases comprising the peptide and steroid therapy in the above formulation.
The peptide lotion therapy is effective for stable vitiligo or segmental vitiligo, but not for fast-diffusing vitiligo.
Steroid therapy is the only effective treatment among the currently known methods of treating fast-diffusing vitiligo (Kendel E, Vitiligo response to 02% betamethasone 17-valerate in flexible collodin “02 in Flexible Collodion % Betamethasone 17-white spots in response to valerate ", Dermetological, 1970, 141, 277-281). However, prolonged treatment with steroids produces many side effects (Geraldiz CB, Gutierrez GT, A clinical trail of clobetasol propionate in Filipino vitiligopatients, Philippine Vitaligopatients Clinical Trials in Clobetasol Propionate Clinical Trials) Clinical Therapy, 1987, 9, 474-482, Ortone J, Clinical Dermato 1987, 7, 120-135). In addition, the disease will recur after discontinuation of treatment. The mode of action of steroids that stop and re-pigment vitiligo is thought to be due to its inhibitory effect on the production of autoantibodies against melanocytes, which has been shown to be responsible for the rapid spread of vitiligo (Han K, Chen D, Bystryn JC, Systemic steroids suppressive lonelocyte antibodies in vitroligo "systemic steroids suppress anti-melanocyte antibodies in vitiligo", J. Cutan. Med. Therefore, it was considered that the combination therapy for vitiligo treatment using topical application of bFGF peptide lotion with oral steroid could reach the following two goals.
1. 1. Stop the rapid spread of vitiligo Faster repigmentation of vitiligo patches.

As a typical example, the efficacy of the peptide in re-pigmentation of vitiligo in human volunteers (FIGS. 1 (a) and (b)) was expressed. FIG. 1 (a) shows the size of vitiligo before the start of treatment with the peptide lotion, and FIG. 1 (b) shows the same vitiligo after topical application of the peptide lotion for 6 months. . However, the treatment was effective in the case of 80% of stable vitiligo and segmented vitiligo (see Tables 1 and 2 for more details).

  Applicable patents describing the peptides for use as pigmented drugs include US Pat. No. 6,143,723, Australian Patent No. 722626, Indian Patent Nos. 185613, 186437 and 186703.

Topical application of the peptide in the above formulation is effective in more than 80% of stable generalized or segmental vitiligo. Even better results with repigmentation rates and success rates of 80% or more appear to be achieved when combined with traditional therapies such as PUV-A therapy for vitiligo. In addition, the peptide lotion therapy is not effective in preventing the rapid spread of vitiligo, and perhaps synergistic combination therapy is traditionally used in cases of fast diffuse vitiligo It appears when used in combination with steroid therapy. Similarly, topical application of the peptide lotion therapy is advantageously used with any other therapy, including surgical therapy.

The mode of action of steroids that stop and re-pigment vitiligo is thought to be due to its inhibitory effect on the production of autoantibodies against melanocytes, which has been shown to be responsible for the rapid spread of vitiligo (Han K, Chen D, Bystryn JC, Systemic steroids suppressive lonelocyte antibodies in vitroligo "systemic steroids suppress anti-melanocyte antibodies in vitiligo", J. Cutan. Med. Therefore, it was considered that the combination therapy for the treatment of vitiligo using the topical application of the peptide lotion with oral steroid could reach the following two goals.
3. 3. Stop the rapid spread of vitiligo Faster repigmentation of vitiligo patches.

This has been shown to occur in clinical trials with human volunteers. The above pair of goals has been achieved. A typical case of this treatment is shown in FIGS. 2 (a) and 2 (b). FIG. 2 (a) describes two vitiligo at the start of oral steroid treatment, and (b) at the end of 3 months of treatment with oral steroids, at the same time the peptide lotion was administered to oral steroids. During the duration of ingestion, the lower vitiligo describes the same vitiligo during topical application once a day. It can be clearly seen that there was a faster re-pigmentation when the peptide lotion was applied compared to the upper vitiligo where the peptide lotion was not applied. A fast cessation of the disease was achieved to 88%, similar to the end of 6 months of treatment with oral steroids, and the results were not altered by topical application of the peptide lotion (more specifically) See Table 3).

At the end of the three-month combination treatment with topical application of steroids and the peptide lotion, more than three times more patients developed pigmentation to a much greater extent than when using steroids alone (more details 4). At the end of 3 months of treatment, the proportion of volunteers who achieved moderate and marked re-pigmentation with combination therapy was double that of steroid therapy alone.

A synergistic combination therapy for the treatment of stable vitiligo comprising the peptide and oral psoralen plus UV-A application in the above formulation.
The main axis or mainstay of stable vitiligo treatment around the world is currently psoralen and UV-A therapy (Mofty EL, A preliminary clinical report on the treatment of leukoderma "Preliminary clinical report in the treatment of vitiligo", with ami in Maj. Egypt.Med.Assoc, 1948, 651-660, Mofty EL, In Villiligo, and psoralens “In Vitiligo and Psoralen”, 1968, Pregman Press Oxford, Ortonne JP, Boseh, White Where are we? ", Pig. Cell. Res 1993, 6, 61-72). However, the effectiveness is limited to partial (Westerof Wiete, Ludmila Nieuwebor-Krobotova, Treatment of vitality with UV-B radiation vs. UV-A white spot UV-A white spot -Treatment with A ", arch. Dermatol 1997, 1525-1528), requiring long duration treatment. In addition, there are a number of side effects, particularly in the liver, and in cancer risk in the Caucasian skin types II and III populations (Henseleer T, Christopher E, Honningsmann H, Wolff K, Skin tumors in European PUV- A study, 8 year follow up of 1643 patients treated with PUV-A for psoriasis, “8 years follow-up of 1643 patients treated with PUV-A for skin tumors, psoriasis in European PUV-A studies”, J. Am. Acad. Dermatol 1997, 16, 108-116, Stern RS, Lard M, The Carcinogenic Risk of Treatment of Treatment. r. So in fact,
a) reduce the duration of treatment,
b) increase the success rate,
c) treating cases that do not respond to PUV-A therapy;
It is necessary.

  The mechanism of action of psoralen and UV-A vitiligo repigmentation remains a mystery. Psoralen itself is not effective in the treatment of vitiligo without subsequent exposure of the skin to UV-A (Gupta AK, Anderson TF, Psoralen photochemotherapy “Soralen photochemotherapy”, J. Amer. Acad. Dermatol 1987, 17). , 703-34). The absorption maximum of psoralen is in the range of 210-330 nm (Harbar LC, Bickers. DR, “Photosensitivity Diseases” “Dawn disease”, 1st edition Philadelphia: WB Saunders 1981, PP 45-55). The action spectrum is probably in the range of 320-335 nm (Flemming MG, Brody N, Biological effective of black light fluorescent lamps for PUV-A “Biological efficacy of black light fluorescent lamps against PUV-A”, J. Amer. Acad. Dermatol, 1985, 12, 894). Psoralen induces skin photosensitization, involves psoralen interchelation with cellular DNA, and ultimately leads to death of the cell. This was also true for melanocytes, as shown by exposure of melanocytes in culture to PUV-A (KaoC, Yu H, Comparison of the effect 8-methoxy psallen plus UV-A on human melanocytes in). vitiligo vulgaris in-vitro "Comparison of in vitro effects of human melanocytes in normal vitiligo with 8-methoxypsoralen + UV-A", J. Invest. Dermatol. 1992, 98, 734-40). And how to explain the beneficial effects of PUV-A in increasing the proliferation of melanocytes, their movement to vitiligo and increasing such repigmented vitiligo? The main target of PUV-A is the epidermis, which receives UV-A radiation. This causes the keratinocytes of the epidermis to die. And the cell contents of keratinocytes leak and stimulate proliferation of adjacent melanocytes. The cell content of keratinocytes contains many mitogens for melanocytes in addition to bFGF (Halban R, Tyrrel L, Longly J, Yanden Y, Rubin J, Ann. NY. Acad. Sci 1993, 680, 290-). 300). It was shown in 1989 that sera from vitiligo patients who were successfully treated with PUV-A stimulate the proliferation of melanocytes from normal untreated vitiligo patients (Ramaiah A, Puri. N, Mojamdar M, A new hypothesis for the etiology of vitality, “a new hypothesis about the etiology of vitiligo”, Acta Derm, Venerol (Stockholm), 1989, 69, 323-327).

  In addition, melanocytes from vitiligo patients who have been treated successfully grow in less generation time than melanocytes from untreated normal individuals (Ramaiah. A, Puri. N, Mojmadar M, A new hyperthesis of the ethology of vitality “ New hypothesis on the pathogenesis of vitiligo ", Acta Derm, Venerol (Stockholm), 1989, 69, 323-327).

  These results show that combined treatment of vitiligo including topical application of the peptide lotion with oral psoralen + UV-A works synergistically and produces faster repigmentation than either of them alone. It was.

3 (a), (b) and FIGS. 4 (a), (b) that the combined application for topical application of the peptide lotion agent and treatment of vitiligo including PUV-A actually works synergistically. It can be seen in the typical photographic diagram shown. FIG. 3 (a) and FIG. 4 (a) are photographs of two volunteers before the combination therapy, and FIG. 3 (b) and FIG. 4 (b) are three months of combination therapy for the same volunteer, respectively. In the photographic diagram at the end of, additional treatment was performed on the right hand of each volunteer using the topical application of the peptide lotion. As can be seen from the picture, the right hand of both volunteers treated with topical application of the peptide lotion is the left hand of both volunteers treated with PUV-A therapy alone Many times more re-pigmentation. The data presented in Table 4 shows that for 7 volunteers treated with topical application of the peptide lotion and combination therapy with PUV-A, compared to treatment with PUV-A alone. It clearly shows that there was significant repigmentation during the three months. The proportion of volunteers with moderate to outstanding rates of repigmentation at the end of 3 months was about twice that of PUV-A alone (see Table 5 for details). See).

Synergistic combination therapy for the treatment of vitiligo that does not respond to PUV-A, including topical application of the peptide and PUV-A therapy.
In response to PUV-A therapy, less than 20% of vitiligo is fully repigmented and 30-40% of vitiligo is partially repigmented (Westerhof Wiete, Ludmila Nieuwebor-Krotova, Treatment of vitality with UV- Bradition VS topical positive Plus UV-A "treatment of vitiligo with UV-B radiation versus typical psoralen + UV-A", arch. Dermatol 1997, 1525-1528). For the vitiligo of volunteers who did not respond to PUV-A therapy even after prolonged treatment for more than one year, a combination therapy of topical application of the agonist peptide lotion with continued PUV-A therapy was selected. It was.

The combination therapy was very successful in re-pigmentation within 3 months for vitiligo that had not responded to PUV-A therapy for over a year. Typical results are shown in FIGS. 5 (a), (b) and FIGS. 6 (a), (b) and FIGS. 7 (a), (b). In fact, the repigmentation rate was significant in FIGS. 5 (a), (b) and FIGS. 6 (a), (b). It is also noteworthy that palm vitiligo, which usually does not respond to PUV-A therapy even after an extended duration, responded well to the peptide lotion and PUV-A combination therapy. Responses to combination therapy were shown in more than 90% of cases (see Table 6 for details).

The results presented here are not exhaustive. Combination therapy using topical application of the peptide in the above formulation is for the treatment of various types of vitiligo with a synergistic effect similar to that observed in the case described above, and most of the currently marketed products It can be applied to any other therapy. The peptide in the formulation can be applied topically not only in the form of a lotion, but also in any other form such as gel / ointment / cream, with similar results.

A synergistic combination therapy for the treatment of vitiligo, comprising a surgical application of punch graft / or other topical application of the peptide following other surgical procedures.
The surgical procedure described above provides a last resort for vitiligo treatment. Such treatments include slush transplantation, expansion in culture of autologous melanocytes and its application to cutaneous excised vitiligo, autogenous expansion of keratinocytes and melanocytes in the form of multi-cell layer sheets in culture and their skin excision Application to the isolated vitiligo, or a stamper transplant to the skin-excised vitiligo of pigmented autologous skin. In all these cases, after the surgical procedure, the vitiligo was surgically bandaged with appropriate antibacterial measures for wound healing. The wound heals in a week or so. Thereafter, topical application of the peptide in the above formulation was carried out for 3 months. The treatment results in faster re-pigmentation than usual.

  Photo FIGS. 8 (a) and (b) show the synergistic effect of increasing the rate of repigmentation in (b) where topical application of the peptide in the formulation was followed by a surgical procedure for stamper implantation. Is described.

(A) Vitiligo display prior to treatment with topical bFGF peptide lotion. (B) Display of vitiligo after treatment with topical bFGF peptide lotion. (A) Display of fast diffusible vitiligo before treatment with steroid and topical bFGF peptide lotion. (B) Display of fast diffusible vitiligo after treatment with steroid and topical bFGF peptide lotion. (A) Display of vitiligo before treatment with topical bFGF peptide lotion and PUVASOL. (B) Vitiligo display after treatment with topical bFGF peptide lotion and PUVASOL. (A) Vitiligo display prior to treatment with topical bFGF peptide lotion and PUVASOL. (B) Vitiligo display after treatment with topical bFGF peptide lotion and PUVASOL. (A) Display of vitiligo resistant to PUV-A for over 1 year prior to treatment with topical bFGF peptide lotion and PUV-A. (B) Display of vitiligo resistant to PUV-A for over 1 year after treatment with topical bFGF peptide lotion and PUV-A. (A) Display of vitiligo resistant to PUV-A for over 1 year prior to treatment with topical bFGF peptide lotion and PUV-A. (B) Display of vitiligo resistant to PUV-A for over 1 year after treatment with topical bFGF peptide lotion and PUV-A. (A) Display of vitiligo resistant to PUV-A for over 1 year prior to treatment with topical bFGF peptide lotion and PUV-A. (B) Display of vitiligo resistant to PUV-A for over 1 year after treatment with topical bFGF peptide lotion and PUV-A. (A) The left hand is a vitiligo display after the stamper transplantation but before the start of treatment with the bFGF peptide lotion. The right hand is the control. (B) The left hand shows vitiligo after 2 months of treatment with a bFGF peptide lotion after stamping device transplantation. The right hand is the control.

Claims (4)

A composition for combination therapy for the treatment of disseminated and vitiligo, for use in topical application of an effective amount of the composition comprising:
0.02-5% w / w of bFGF amino acids 106-115 (SEQ ID NO: 1), bFGF amino acids 106-120 (SEQ ID NO: 5), bFGF amino acids 1-24 (SEQ ID NO: 6) and SEQ ID NOs: 2, 3, At least one peptide selected from the group consisting of 4, 7 and 8;
10-50% w / w solvent,
10-40% w / w glycol, and 10-40% w / w at least one penetration enhancer ,
The composition is administered with psoralen and UV-A (PUV-A therapy) / PUVASOL,
The above composition. A composition for combination therapy for the treatment of vitiligo that does not respond to PUV-A therapy for use in topical application of an effective amount of the composition comprising:
0.02-5% w / w of bFGF amino acids 106-115 (SEQ ID NO: 1), bFGF amino acids 106-120 (SEQ ID NO: 5), bFGF amino acids 1-24 (SEQ ID NO: 6) and SEQ ID NOs: 2, 3, At least one peptide selected from the group consisting of 4, 7 and 8;
10-50% w / w solvent,
10-40% w / w glycol, and 10-40% w / w at least one penetration enhancer ,
The composition is administered with PUV-A therapy;
The above composition. A composition for combination therapy for the treatment of fast-diffusing vitiligo for use in topical application of an effective amount of the composition comprising:
0.02-5% w / w of bFGF amino acids 106-115 (SEQ ID NO: 1), bFGF amino acids 106-120 (SEQ ID NO: 5), bFGF amino acids 1-24 (SEQ ID NO: 6) and SEQ ID NOs: 2, 3, At least one peptide selected from the group consisting of 4, 7 and 8;
10-50% w / w solvent,
10-40% w / w glycol, and 10-40% w / w at least one penetration enhancer ,
The composition is administered with steroid therapy;
The above composition. A composition for combination therapy for the treatment of vitiligo for use in topical application of an effective amount of the composition comprising:
0.02-5% w / w of bFGF amino acids 106-115 (SEQ ID NO: 1), bFGF amino acids 106-120 (SEQ ID NO: 5), bFGF amino acids 1-24 (SEQ ID NO: 6) and SEQ ID NOs: 2, 3, At least one peptide selected from the group consisting of 4, 7 and 8;
10-50% w / w solvent,
10-40% w / w glycol, and 10-40% w / w at least one penetration enhancer ,
The composition is administered with a surgical procedure;
The above composition.