Use of α-(8-quinolinyloxy) mono-substituted phthalocyanine zinc for treatment of psoriasis

Abstract

The present invention relates to a use of alpha-(8-quinolinyloxy) mono-substituted phthalocyanine zinc for the treatment of psoriasis. The use is suitable for various type of psoriasis by utilizing photodynamic therapy. The photosensitizer is used to treat psoriasis by utilizing laser wave length of 670 nm, with high light sensitivity, fast photobleaching, short time needed in protection from light, and avoiding potential toxic and side-effect of photochemistry therapy by using ultraviolet irradiation. The preparation used in the use of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc for the treatment of psoriasis includes solution, cream, nanomicelle, microsphere etc, and mode of administration can be systemic or topical administration. The practicability of treating psoriasis by photodynamic therapy has been validated on cell and animal model. The use achieves perfect effect.

Claims

1. A method of treating psoriasis in a patient, wherein the method comprises administering a photosensitizer composition comprising a zinc phthalocyanine compound of the structure: ##STR00001##

2. The method of claim 1, wherein the psoriasis is selected from the group consisting of psoriasis vulgaris, pustula psoriasis, erythrodermic psoriasis or arthritic psoriasis.

3. The method of claim 1, wherein the method of treating comprise a photodynamic therapy using a semiconductor laser with a wavelength of 670 nm as a light source.

4. The method of claim 1, wherein the composition is formulated in a form selected from the group consisting of a solution, a cream, a nanomicelle, a microsphere or a patch.

5. The method of claim 1, wherein the psoriasis is selected from the group consisting of blood heat type psoriasis, blood dryness type psoriasis, or blood stasis type psoriasis.

6. The method of claim 1, wherein the method of treatment comprises a photodynamic therapy using a semiconductor laser.

7. The method of claim 1, wherein the method of treatment comprises a photodynamic therapy using a light source.

8. The method of claim 1, wherein the mode of administration is selected from the group consisting of a systemic administration or a topical administration.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1: Mouse vaginal epithelial basal cells and mitotic cells.

(2) FIG. 2: Effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on the diethylstilbestrol-induced excessive proliferation of mouse vaginal epithelial cells. HE staining. A, blank control group (200); B, model control group (200); C, irradiation without administration group (200); D, PDT group 1 (200); E, PDT group 2 (200) and F, PDT group 3 (200).

(3) FIG. 3: Propranolol-induced psoriasis-like lesions on guinea pig ears. A, normal guinea pig ear. B, model guinea pig ear; changes such as scales, dilatation of blood vessels, thickening of skin can be observed. C, acanthosis (HE staining, 40). D, hyperkeratosis accompanied with parakeratosis (HE staining, 200).

(4) FIG. 4A: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of blank control group.

(5) FIG. 4B: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of model control group.

(6) FIG. 4C: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of irradiation without administration group.

(7) FIG. 4D: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of administration without irradiation group.

(8) FIG. 4E: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of injection administration group.

(9) FIG. 4F: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of topic administration group 1.

(10) FIG. 4G: Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pigs (HE staining, 40) of topic administration group 2.

BEST MODE OF THE INVENTION

(11) 1. Effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on the survival rate of HaCaT cells.

(12) HaCaT cell line, a cell line of normally mutated and immortalized human keratinocyte, is one of the most wildly used cell models for the currently study of psoriasis. As compared with primary cultured keratinocytes, HaCaT cells have similar biological properties, but its operation is simpler.

(13) HaCaT cells were routinely cultured in RPMI-1640 medium containing 10% fetal bovine serum at 37 C. in a 5% CO.sub.2 incubator. The cells were harvested in logarithmic growth phase, digested with 0.25% trypsin (containing EDTA), adjusted to a cell density of 210.sup.5 cells/mL and seeded in a 96-well plate, 100 L per each well; after incubation for 24 hours, -(8-quinolinyloxy) mono-substituted phthalocyanine zinc was added at concentrations of 0.001 g/mL, 0.01 g/mL, 0.1 g/mL, 1 g/mL and 10 g/mL respectively and three complex wells were set for each concentration. A blank control group and a solvent control group were set additionally. After administration, the cells were incubated for additional 4 hours and irradiated by using a photodynamic therapeutic instrument at 670 nm. Irritation conditions: output power, 1500 mW; spot diameter, 8 cm; irradiation time, 180 s. After irradiation, the 96-well plate was placed back to the CO.sub.2 incubator and incubated for additional 48 hours. The culture medium was discarded; 200 L of MTT solution (0.5 mg/mL) was added to each well; the cells were incubated for additional 4 hours; then, 200 L of dimethyl sulfoxide was added to each well and the absorbance value of each well was measured by a microplate reader at 545 nm (reference wavelength: 450 nm). 3 times of parallel measurements were conducted and the mean value was calculated. The inhibition rate of proliferation was calculated and IC50 was calculated by Bliss method.

(14) Results: -(8-quinolinyloxy) mono-substituted phthalocyanine zinc can inhibit the proliferation of HaCaT cells in a concentration-dependent manner, and under the adopted irradiation conditions, the IC50 thereof was 0.1970.022 g/mL.

(15) 2. Effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on the diethylstilbestrol-induced excessive proliferation of mouse vaginal epithelial cells.

(16) Estrogen (for example diethylstilbestrol) may induce the active proliferation of mouse vaginal epithelial cells, thereby accelerating the turnover of epithelial cells; and colchicine may make the cell cycle stagnated in the mitotic metaphase and then the inhibition effects of a test substance on mitosis and excessive cell proliferation may be observed. This model may simulate the characteristic of overly rapid proliferation of epithelial cells in psoriasis patients, is simple and easy to obtain, and is one of the most common animal models in the assessment of drug efficacy for psoriasis.

(17) Sixty ICR mice were taken and randomly divided into 6 groups according to weights: blank control group, model control group, irradiation without administration group and PDT groups 1, 2 and 3. Except for the blank control group, all of other groups were intraperitoneally injected with diethylstilbestrol (0.2 mg/mouse), once-daily for 3 days. At the 4.sup.th day, PDT groups 1, 2 and 3 were intraperitoneally injected with -(8-quinolinyloxy) mono-substituted phthalocyanine zinc solution at doses of 1.2 mg/kg, 0.6 mg/kg and 0.3 mg/kg, respectively; and other 3 groups were not administrated. After protection from light for 6 hours, the PDT groups 1, 2 and 3 and the irradiation without administration group were subjected to laser irradiation by using a semiconductor photodynamic therapeutic instrument; the irradiation conditions were 100 mW540 s, 300 mW60 s, 100 mW60 s and 300 mW540 s, respectively; the spot diameter was 3 cm; and the abdomen of mice was irradiated. The blank control group and the model control group were not subjected to irradiation. After one week post-irradiation, mice of each group were intraperitoneally injected with colchicine at a dose of 2 mg/kg, making the cell mitosis stagnated in the mitotic metaphase. After 4 hours post-injection of colchicine, animals were sacrificed by anesthesia. Mouse vagina was taken, fixed in 10% formaldehyde solution, conventionally dehydrated, embedded in paraffin, sliced and stained with HE; 300 basal cells were counted under a microscope and the number of mitotic cells was calculated and the mitotic index was calculated (number of mitotic cells per 100 basal cells).

(18) Results: diethylstilbestrol could induce the excessive proliferation of ICR mouse vaginal epithelial cells; as compared with the blank control group, the mitotic index of the model control group was increased significantly; -(8-quinolinyloxy) mono-substituted phthalocyanine zinc-PDT could inhibit the excessive proliferation of mouse vaginal epithelial cells; as compared with the model control group, the mitotic index of PDT groups 1, 2 and 3 was significantly reduced. For details, see Table 1 and FIGS. 1-2.

(19) TABLE-US-00001 TABLE 1 Effects of -(8-quinolinyloxy) mono-substituted phthalocyanine-PDT on excessive proliferation of mouse vaginal epithelial cells (x s, n = 10). Irradiation Output Dose Time power Mitotic Index Group No. (mg/kg) (s) (mW) (%) Blank control group 0 0 0 5.3 0.9.sup.## Model control group 0 0 0 19.5 6.7** Irradiation without 0 540 300 13.7 5.3*.sup.## administration group PDT group 1 2 540 100 6.4 2.2.sup.##{circumflex over ()} PDT group 2 1 60 300 7.2 2.6.sup.## PDT group 3 0.5 60 100 10.3 4.3*.sup.## Note: as compared with the blank control group, *p < 0.05, **p < 0.01; as compared with the model control group, .sup.##p < 0.01; and as compared with the irradiation without administration group, {circumflex over ()}p < 0.05.

(20) 3. Therapeutic effect of -(8-quinolinyloxy) mono-substituted phthalocyanine zinc on propranolol-induced psoriasis-like lesions in guinea pig.

(21) Propranolol is a -adrenergic receptor blocking agent, and it may be applied to guinea pig skin to block the -adrenergic receptor in keratinocytes and thereby reducing the intracellular cAMP level, leading to histopathological changes similar to psoriasis such as hyperkeratosis, parakeratosis and acanthosis of animal epidermis, which are the same as human psoriatic lesions at the molecular level. Application of propranolol to ear skin of guinea pig to replicate a psoriasis-like skin model is often used for observing the therapeutic effect of a drug on psoriasis.

(22) Preparation method for 5% propranolol liniment: dissolving 5 g powder of propranolol API in 50 mL of 50% ethanol, adding 5 g of polyoxyethylene pyrrolidone 30, 2.5 mL of azone, 2.5 mL of propylene glycol, and finally adding 50% ethanol to 100 mL. 5% propranolol was prepared fresh for use. Forty-two guinea pigs were randomly divided into a blank control group and a model-establishing group according to weights, 6 guinea pigs in the blank control group and 36 guinea pigs in the model-establishing group. 5% propranolol was applied uniformly on the back of both ears of guinea pigs in the model-establishing group at a dose of 0.1 mL/ear, once in the morning and once in the afternoon, and the administration lasted for continuous 4 weeks. After the model was established, the guinea pigs in the model-establishing group were randomly divided into 6 groups according to weights again, 6 guinea pigs in each group. The 6 groups were model control group, irradiation without administration group, administration without irradiation group, injection administration group and topical administration groups 1 and 2, respectively. The administration without irradiation group and the injection administration group were intravenously injected with -(8-quinolinyloxy) mono-substituted phthalocyanine zinc solution at a dose of 0.45 mg/kg and the topical administration groups 1 and 2 were respectively applied with 5% and 1% -(8-quinolinyloxy) mono-substituted phthalocyanine zinc liniments at a dose of 0.1 mL/ear. Other 3 groups were not administrated. After protection from light for 24 hours, the irradiation without administration group, the injection administration group and the topical administration groups 1 and 2 were subjected to laser irradiation by using a semiconductor photodynamic therapeutic instrument with the following irradiation conditions: output power, 1,000 mW; spot diameter, 6 cm; irradiation time, 399 s; irradiation dose, 14 J/cm.sup.2. After irradiation, animals were protected from light for 48 hours, and then recovered for 3 days. A photodynamic therapy (administration, irradiation, protection from light and recovery) was repeated once according to the abovementioned steps. After the therapy, animals of each group were sacrificed by anesthesia. Skin of both sides of ear was fixed in 10% formaldehyde solution, conventionally dehydrated, embedded in paraffin, sliced and stained with HE, and observed under a microscope.

(23) Results: Application of 5% propranolol to skin on the back of ear of guinea pigs for 4 weeks can result in obvious psoriasis-like lesions; thickening of ear skin, dilatation of blood vessels and scaly lesions can be observed with naked eyes; acanthosis, hyperkeratosis and parakeratosis and the like can be observed under a microscope. For details, see FIG. 3. In the irradiation without administration group and the administration without irradiation group, such lesions got improved to some extent but not obviously. In the injection administration group and the topical administration groups 1 and 2, propranolol-induced psoriasis-like lesions of the guinea pig ear were improved significantly; after two photodynamic therapies, the prickle cell layer got thinner significantly, hyperkeratosis and parakeratosis got alleviated; some guinea pig ears were recovered to normal and had no obvious difference as compared with the blank control group. For details, see FIG. 4.

(24) The above examples are merely the preferred examples of the present invention, and variations and modifications that are made according to the scope of the present invention come within the scope of the present invention.