U.S. patent application number 12/317642 was filed with the patent office on 2009-07-16 for compositions for the reversal of stratum granulosum in keratinization disorders.
This patent application is currently assigned to Apptec, Inc.. Invention is credited to N.R. Murugan, N.B. Baktha Reddy, Vilambi Nrk Reddy, Anil Torgalkar.
Application Number | 20090181113 12/317642 |
Document ID | / |
Family ID | 40850843 |
Filed Date | 2009-07-16 |
United States Patent
Application |
20090181113 |
Kind Code |
A1 |
Reddy; N.B. Baktha ; et
al. |
July 16, 2009 |
Compositions for the reversal of stratum granulosum in
keratinization disorders
Abstract
Compositions that, when used topically in a therapeutically
effective amount, are safe and effective for the regression of
stratum granulosum in patients exhibiting keratinization disorders,
such as psoriasis. The compositions, which can be formulated as
ointments, oils, soaps and shampoos, comprise a non-aqueous extract
of Wrightia tinctoria, an extract of Cocos nucifera, and
pharmaceutically or cosmetically acceptable excipients suitable for
topical use.
Inventors: |
Reddy; N.B. Baktha;
(Chennai, IN) ; Reddy; Vilambi Nrk; (Trichy,
IN) ; Torgalkar; Anil; (Crambury, NJ) ;
Murugan; N.R.; (Trichy, IN) |
Correspondence
Address: |
BREGEN TECHNICAL CONSULTANTS L.L.C.
154 OLD CLINTON ROAD
FLEMINGTON
NJ
08822
US
|
Assignee: |
Apptec, Inc.
|
Family ID: |
40850843 |
Appl. No.: |
12/317642 |
Filed: |
December 23, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11731345 |
Mar 31, 2007 |
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12317642 |
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11288923 |
Nov 28, 2005 |
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11731345 |
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Current U.S.
Class: |
424/727 |
Current CPC
Class: |
A61Q 19/00 20130101;
A61P 17/06 20180101; A61P 17/00 20180101; A61Q 5/02 20130101; A61Q
19/10 20130101; A61K 8/9789 20170801; A61K 36/24 20130101; A61K
36/889 20130101; A61K 8/9794 20170801; A61K 36/24 20130101; A61K
2300/00 20130101; A61K 36/889 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/727 |
International
Class: |
A61K 36/889 20060101
A61K036/889; A61P 17/06 20060101 A61P017/06; A61P 17/00 20060101
A61P017/00 |
Claims
1. An herbal composition that, when used topically in
therapeutically effective amounts, is effective for the regression
of stratum granulosum in keratinization disorders, comprising: an
extract of Wrightia tinctoria in a non-aqueous medium, an extract
of Cocos nucifera and pharmaceutically or cosmetically acceptable
excipients for use in ointment, oil, soap and shampoo
formulations.
2. The herbal composition according to claim 1, wherein the
non-aqueous medium for the herbal extract is a non-volatile
oil.
3. The herbal composition according to claim 2, wherein the
non-volatile oil is a vegetable oil, selected from the group:
coconut oil, gingely oil (sesame seed oil), sunflower oil, corn
oil, and a refined vegetable oil.
4. The herbal composition according to claim 2, wherein the
non-volatile oil is present in the extract in an amount of from 80%
to 99% by weight of the extract.
5. The herbal composition according to claim 1, wherein the
Wrightia tinctoria is obtained from at least one of the leaves,
twigs, flowers, and fruit of the Wrightia tinctoria plant.
6. The herbal composition according to claim 5, wherein the herbal
extract of Wrightia tinctoria is present in the non-aqueous medium
in an amount of from 1% to 20% by weight.
7. The herbal composition according to claim 1, wherein the Cocos
nucifera is extracted from the copra of the coconut.
8. The herbal composition according to claim 7, wherein the Cocos
nucifera is present in the amount of 40% to 80% by weight.
9. An ointment formulation for the regression of stratum granulosum
in keratinization disorders, that, when used topically in
therapeutically effective amounts, comprises the herbal composition
according to claim 1 and pharmaceutically acceptable
excipients.
10. An oil formulation for the regression of stratum granulosum in
keratinization disorders, that, when used topically in
therapeutically effective amounts, comprises the herbal composition
according to claim 1 and pharmaceutically acceptable
excipients.
11. A liquid soap formulation for the regression of stratum
granulosum in keratinization disorders, that, when used topically
in therapeutically effective amounts, comprises the herbal
composition according to claim 1 and pharmaceutically acceptable
excipients.
12. A shampoo formulation for the regression of stratum granulosum
in keratinization disorders, that, when used topically in
therapeutically effective amounts, comprises the herbal composition
according to claim 1 and pharmaceutically acceptable
excipients.
13. A method for the regression of stratum granulosum in
keratinization disorders, which comprises administering to a
subject in need thereof a formulation comprising a therapeutically
effective amount of a composition according to claim 1, said
formulation in a form chosen from the group: an oil, a shampoo, an
ointment and a liquid soap.
14. An herbal composition for the regression of stratum granulosum
in psoriatic lesions when used topically in therapeutically
effective amounts, comprising: an extract of Wrightia tinctoria in
a non-aqueous medium, an extract of Cocos nucifera and
pharmaceutically or cosmetically acceptable excipients for use in
ointment, oil, soap and shampoo formulations.
15. The herbal composition according to claim 13, wherein the
non-aqueous medium for the herbal extract is a non-volatile
oil.
16. The herbal composition according to claim 14, wherein the
non-volatile oil is a vegetable oil, selected from the group:
coconut oil (Cocos nucifera), gingely oil (sesame seed oil),
sunflower oil, corn oil, and a refined vegetable oil.
17. The herbal composition according to claim 14, wherein the
non-volatile oil is present in the extract in an amount of from 80%
to 99% by weight of the extract.
18. The herbal composition according to claim 14, wherein the
Wrightia tinctoria is obtained from at least one of the leaves,
twigs, flowers, and fruit of the Wrightia tinctoria plant.
19. The herbal composition according to claim 17, wherein the
herbal extract of Wrightia tinctoria is present in the non-aqueous
medium in an amount of from 1% to 20% by weight.
20. The herbal composition according to claim 18, wherein the Cocos
nucifera is extracted from the copra of the coconut.
21. The herbal composition according to claim 19, wherein the Cocos
nucifera is present in the amount of 40% to 80% by weight.
22. An ointment formulation for the regression of stratum
granulosum in psoriatic lesions that, when used topically in
therapeutically effective amounts, comprises the herbal composition
according to claim 13 and pharmaceutically acceptable
excipients.
23. An oil formulation for the regression of stratum granulosum in
psoriatic lesions that, when used topically in therapeutically
effective amounts, comprises the herbal composition according to
claim 13 and pharmaceutically acceptable excipients.
24. A liquid soap formulation for the regression of stratum
granulosum in psoriatic lesions that, when used topically in
therapeutically effective amounts, comprises the herbal composition
according to claim 13 and pharmaceutically acceptable
excipients.
25. A shampoo formulation for the regression of stratum granulosum
in psoriatic lesions that, when used topically in therapeutically
effective amounts, comprises the herbal composition according to
claim 13 and pharmaceutically acceptable excipients.
26. A method for the regression of stratum granulosum in psoriatic
lesions which comprises administering to a subject in need thereof
a formulation comprising a therapeutically effective amount of at
least one composition according to claim 1, said formulation in a
form chosen from the group: an oil, a shampoo, an ointment and a
liquid soap.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application, filed under 35 U.S.C. 120, is a
continuation-in-part of U.S. non-provisional patent application
Ser. No. 11/731,345 entitled "Compositions for the Reversal of
Stratum Granulosum in Keratinization Disorders" filed on Mar. 31,
2007, and as such claims priority benefit under 35 U.S.C. 120 of
this application. The present application is related to U.S.
non-provisional applications Ser. No. 11/288,923 filed on Nov. 28,
2005; Ser. No. 12/009,799 filed on Jul. 3, 2008; and Ser. No.
12/286,564 files on Sep. 30, 2008. The disclosures of these
applications are incorporated herein by reference in their
entireties.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] Not Applicable
THE NAMES OF THE PARTIES OF A JOINT RESEARCH AGREEMENT
[0003] Not Applicable
INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT
DISC
[0004] Not Applicable
FIELD OF THE INVENTION
[0005] The present invention relates to compositions for the
reversal of stratum granulosum in keratinization disorders. In
particular, the present invention relates to herbal compositions
for the regression and reversal of stratum granulosum in
keratinization disorders in psoriatic lesions.
BRIEF DESCRIPTION OF THE BACKGROUND ART
[0006] The skin is made up of two primary layers that differ in
function, thickness, and strength. From outside to inside, they are
the epidermis and its sublayers, and the dermis, after which is
found subcutaneous tissue, or the hypodermis.
[0007] The epidermis, the outermost layer of skin, is thin but
complex. Melanin, which is responsible for skin pigmentation, is
found throughout the epidermis. The epidermis also keratinizes to
produce nails, hair, sweat, and to regenerate.
[0008] Keratinization, the maturation and migration of skin cells,
begins in the innermost layer of the epidermis, the stratum
germinativum [see item E in FIG. 1]. These cells, called
keratinocytes, accumulate and move outward toward the next
epidermis layer, the stratum spinosum [see item D in FIG. 1], where
they become dense. The next layer, known as the stratum granulosum
[see item C in FIG. 1] layer, contains 1 to 3 rows of flattened
cells whose cytoplasm contain small granules. The granules contain
proteins being transformed into the waterproofing protein keratin.
It is in this layer that one finds glycolipids and a thickening of
the membrane. A protein called filigrin is made in this layer and
is put in the granules. In this layer, cells lose their nuclei. In
the cytoplasm, there are keratohyalin granules as well as
membrane-coating granules which expel their lipid contents into the
intercellular spaces. Lipids assist in the formation of water
barriers among the cells of the skin, which, in turn, help to
ensure body moisturization. At this point, the cell also becomes
flattened, or horny, and the nucleus disappears; what remains is
keratin. In the next layer, the stratum lucidum [see item B in FIG.
1], the cell is prepared to move into its final sublayer with the
addition of melanin granules. Then, sudden changes in enzyme
function cause cell death. The products of this ongoing process
form the stratum corneum [see item A in FIG. 1], which is the
outermost epidural layer consisting of neatly packed dead horny
cells.
[0009] Keratinization disorders in the stratum granulosum layer in
the epidermis can often lead to clinically significant skin
manifestations. One common disorder includes thinning of the
stratum granulosum layer due to malfunctioning of the
keratinization process leading to reduction in the moisture barrier
properties of the stratum granulosum layer. In addition, for
example, over activated keratinocytes actively producing and
secreting pro-angiogenic factors in the form of growth factors or
cytokines can result in increased blood vessel formation in the
papillary dermis which may sometime extend into the epidermis.
Epidermal microvascular proliferation ultimately leads to epidermal
keratinocyte hyperproliferation, thickening of the epidermis with
parakeratosis of the stratum corneum and inflammatory infiltrate
around the blood vessels in the papillary dermis [see FIG. 1]. The
microvascular changes are also characterized by increased
tortuosity of dermal capillary loops which precedes the development
of epidermal hyperplasia. Mitotic activity in the basal and
suprabasal cells are greatly increased [Dr. George Jacob, Seminar
on Psoriasis, Dubai, January 2001]. Cellular invasion takes place,
particularly in the suprapapillary region to form the Munro `micro
abscess` which are extruded in the horny layer or they may collect
in disintegrated malphigian cells, the cytoplasm of which had been
lysed to form the multilocular or stratum granulosum of Kogoj.
[0010] Stratum granulosums of Kogoj are multilocular pustules in
the upper stratum malpighii within a sponge-like network made up of
flattened keratinocytes [M. S. Stone and T. L. Ray, DermPath Tutor,
Department of Dermatology, Iowa College of Medicine, September
1995]. They are seen in psoriasis, Reiter's disease, geographic
tongue and rarely in candidiasis. Histological studies, including
immunocytochemistry, routine histology and electron microscopy have
clearly established that alterations in the blood vessel formation
of the skin discussed above are a prominent feature of psoriasis
and there is a marked increase in cutaneous blood active edge of
the psoriatic plaque [Braverman I M, Yen A. Ultrastructure of the
capillary loops in the dermal papillae of psoriasis. J Invest
Dermatol 1977: 68: 53-60].
[0011] Numerous therapies in the field of allopathy medicine have
been reported in reducing a stratum granulosum disorder, especially
in relation to psoriasis: [0012] Treatment of psoriasis--Part
1--Topical Therapy and Phototherapy, Mark Lebwohl, MD, et al,
American Academy of Dermatology, October 2001 Vol 45 (4). [0013]
Treatment of psoriasis--Part 2--Systemic Therapies, Mark Lebwohl,
MD, et al, American Academy of Dermatology, November 2001 Vol
45(5). [0014] The immunological basis for the treatment of
psoriasis with new biological agents. James. G. Krueger, M.D,
American Academy of Dermatology, June 2002 Vol 46(1) Pages 1-26.
[0015] New psoriasis treatments based upon a deeper understanding
of the pathogenesis of psoriasis vulgaris and psoriatic arthritis.
Jeffrey P. Callen et al, American Academy of Dermatology, August
2003 Vol 49(5) Pages 351-356.
[0016] However, most of these therapies provide only temporary
symptomatic relief and are either unsatisfactory or very expensive
and are associated with either short term or long term undesired
side effect profiles. [National Psoriasis Conference, Boston Plaza
Hotel, Aug. 5-8, 2005, Boston, Mass., USA.] In addition, it is
possible for the treatments to lessen the severity of one or more
of the manifestations of psoriasis, but not all. These
manifestations include but are not limited to stratum granulosum of
Kogoj, dermal vessel tortuosity, and spongiform pustules.
[0017] It is well known that herbal formulations often have fewer
undesirable side effect profiles and hence provide a viable
alternative therapy to manage the keratinization disorders that
exhibit stratum granulosum. Research efforts to develop herbal
formulations to treat these conditions have been on the rise and
there is a continuing need to develop herbal formulations to treat
stratum granulosum in keratinization disorders with minimal or no
side effects: [0018] Chopra, R. N., Nayar, S. C., and Chopra I. C.,
Glossary of Indian Medicinal Plants, C.S.I.R., P.259, 1956. [0019]
Murugesa Mudaliar, K. S., Gunapadam (Material Medica) Vegetable
Section, Govt. of TamilNadu, P. 527 (1969). [0020] Venkatarajan,
S., Sarabendra Vaithiya Muraigal, P. 160, 161 & 167 (1965).
[0021] Wealth of India, Raw Materials, Vol. X, P. 588-590, CSIR.,
New Delhi (1976). [0022] Yugimuni Vaidya Chintamani (800) Stanza
494-518, B. Rathina Nayakar & Sons, Madras, India. [0023] Nair,
C. P. R., Kurup, P. B., Pillai, K. G. B., Geetha, A., and Ramiah,
N., Effect of Nimbidin in Psoriasis, Indian Medical Journal,
October 1978.
[0024] The invention provides herbal formulations, developed by a
dermatologist, based on Wrightia tinctoria and an extract of Cocos
nucifera which, when used as directed in therapeutically effective
amounts, have been clinically proven to be safe and efficacious in
reversing or regressing stratum granulosum in keratinization
disorders in humans in need of treatment. The definition of the
term herb, as it is used here is taken from the definition provided
by the Herb Society of America: The tem herb refers to a wide range
of plants, including perennials, trees, shrubs, annuals, vines, and
more primitive plants, such as ferns, mosses, algae, lichens, and
fungi. The herbs are valued for their flavor, fragrance, medicinal
and healthful qualities, economic and industrial uses, pesticidal
properties, and coloring materials (dyes)." [Bown, Deni. The Herb
Society of America New encyclopedia of Herbs and Their Uses. New
York: Dorling Kindersley, 2001, p. 18]
SUMMARY OF THE INVENTION
[0025] It is an object of the present invention to provide a
composition that, when used topically as directed, is suitable for
the reversal of stratum granulosum in keratinization disorders. The
composition can be formulated, for example, as an ointment, an oil,
a soap or a shampoo. The composition is comprised of efficacious
amounts of a non-aqueous extract of Wrightia tinctoria, an extract
of Cocos nucifera and suitable pharmaceutically or cosmetically
acceptable excipients designated for topical use in humans.
BRIEF DESCRIPTION OF THE DRAWINGS
[0026] FIG. 1: Illustration of the stratum granulosum layer in the
epidermis
[0027] FIG. 2: Micrographs illustrating stratum granulosum in
keratinization disorders--before and after treatment
DETAILED DESCRIPTION OF THE INVENTION
[0028] The present invention relates to formulations, which
unexpectedly provide statistically superior efficacy to allopathy
control formulations in reversal of stratum granulosum disorder
when used in therapeutically effective amounts. They are proven
safe to use in humans when used topically as directed.
[0029] It is an object of the invention to provide a composition
that comprises a non-aqueous extract of Wrightia tinctoria, an
extract of Cocos nucifera, and pharmaceutically or cosmetically
acceptable excipients suitable for topical use. The composition can
be formulated as an ointment, an oil, a soap and a shampoo, and,
when a therapeutic amount is applied topically to the affected
area, in the treatment of stratum granulosum in keratinization
disorders and is safe in humans.
[0030] The herbal extract in the topical composition for the
regression of stratum granulosum in keratinization disorders is
derived from the Wrightia tinctoria plant, especially the leaves,
twigs, flowers, fruits or a combination of these parts of the
plant. Wrightia tinctoria is an apocynaceae tree growing throughout
India. Its flowers are white and fragrant.
[0031] It is another object of the invention to provide a process
for preparing the non-aqueous extract of Wrightia tinctoria. The
non-aqueous extract is prepared at ambient temperature by cleaning
and pulverizing the selected parts of the Wrightia tinctoria plant
and soaking them in a non-aqueous oil medium containing coconut
oil. Care should be taken to add sufficient oil medium to ensure
that the plant material is completely submerged. The plant
material/oil medium is then irradiated with a light source in the
spectrum range of 300-1100 nm for a period of approximately 5 days.
During this time the herbal ingredients are allowed to extract into
the non-aqueous oil medium. At the end of the extraction, the oil
medium is a purplish brown color. It is then filtered and the
filtrate is stored for further processing as the non-aqueous herbal
extract of Wrightia tinctoria.
[0032] Other herbal extracts may optionally be included in the
formulation, including Melia azadirachta Linn oil (Neem oil), which
has been documented to have beneficial skin effects [Nair et al.,
1978]. The topical composition of the invention for the reversal of
stratum granulosum in keratinization disorders optionally comprises
an extract of the active herbal ingredient mentioned above in the
extraction medium in the amount from 1% to 20% by weight.
[0033] The herbal extract of Cocos nucifera in the composition of
this invention is derived from the copra of the coconut. The copra
of the coconut is dried and then processed by grinding and pressing
to extract the oil, which is then purified and stabilized. The
composition of the present invention comprises the herbal extract
(the oil) of Cocos nucifera present in the amount of 40% to 80% by
weight.
[0034] It is an object of the invention to provide formulations for
topical use by further compounding the compositions of the
invention with ingredients mentioned herein to prepare
formulations, including but not limited to, an ointment, an oil, a
liquid soap and a shampoo.
[0035] The ointment formulation of the herbal composition of the
invention suitable, when used topically in a therapeutically
effective amount, for the reversal of stratum granulosum in
keratinization disorders, includes pharmaceutically acceptable
excipients such as beeswax, paraffin (liquid, soft and hard), and
other standard ointment bases or their equivalents to optimize the
use characteristics of the formulations, such as consistency and
spreadability, as well as manufacturability and stability. The
ointment composition comprises one or more of the excipients
including beeswax, optimally present in the amount of 1 to 5% by
weight; paraffin, optimally present in the amount of 5 to 40% by
weight; and standard ointment bases, optimally present in the
amount of 5 to 50% by weight.
[0036] The oil formulation of the herbal composition of the
invention suitable, when used topically in a therapeutically
effective amount, for the reversal of stratum granulosum in
keratinization disorders, includes pharmaceutically acceptable
excipients such as vegetable oil, animal oil, and synthetic oils
such as mineral oil and liquid paraffin or their equivalents to
optimize the use characteristics of the formulations, such as
consistency and spreadability, as well as manufacturability and
stability. Preferentially, the oil composition comprises
excipients, such as coconut oil, present in the amount of 70 to 95%
by weight.
[0037] The liquid soap formulation of the herbal composition of the
invention suitable, when used topically in a therapeutically
effective amount, for the reversal of stratum granulosum in
keratinization disorders, includes pharmaceutically acceptable
excipients, including but not limited to: water, surface active
agents, thickeners and viscosity enhancers, foam boosters, and
stabilizers to optimize the use characteristics, such as
consistency, cleaning, spreadability and foaming, as well as
manufacturability and stability. The liquid soap formulation of the
present invention preferentially comprises excipients such as water
present in the amount of 60 to 85% by weight; surface active agents
present in the amount of 5 to 40% by weight; thickeners or
viscosity enhancers present in the amount of 0.5 to 8% by weight;
foam boosters present in the amount of 1 to 4% by weight; and
stabilizers present in the amount of 0.5 to 2% by weight.
[0038] The shampoo formulation of the herbal composition of the
invention suitable, when used topically in a therapeutically
effective amount, for the reversal of stratum granulosum in
keratinization disorders, includes pharmaceutically acceptable
excipients, comprising water, surface active agents, thickeners or
viscosity enhancers, foam boosters, and stabilizers to optimize the
use characteristics such as consistency, cleaning, spreadability
and foaming, as well as manufacturability and stability. The
shampoo composition of the present invention preferentially
comprises excipients including water present in the amount of 50 to
85% by weight; surface active agents present in the amount of 10 to
30% by weight; thickeners or viscosity enhancers present in the
amount of 2 to 8% by weight; foam boosters present in the amount of
2 to 6% by weight; and stabilizers present in the amount of 0.5 to
2% by weight.
[0039] In addition, the ointment, oil, liquid soap and shampoo
formulations of the herbal composition of the invention suitable,
when used topically in a therapeutically effective amount, for the
reversal of stratum granulosum in keratinization disorders,
optionally comprise preservatives, coloring agents and fragrances
as needed, wherein the preservatives, coloring agents and
fragrances are present in the amount of 0 to 5 total weight %.
[0040] Safety and efficacy studies were conducted on subjects
exhibiting of stratum granulosum in keratinization disorders using
topical formulations of the herbal composition of the present
invention described above, containing a non-aqueous herbal extract
of Wrightia tinctoria, an herbal extract of Cocos nucifera and
pharmaceutically or cosmetically acceptable excipients. Patients
suffering from chronic inflammatory skin conditions selected for
the study exhibited stratum granulosum in keratinization disorders
in the form of psoriatic lesions. Psoriasis is a representative
example of a condition exhibiting stratum granulosum in
keratinization disorders. The results are illustrated by the
following example.
EXAMPLE
[0041] Twenty patients were enrolled in a clinical study and were
divided into two groups of 10 patients each. Group I was treated
with the herbal formulation (see Table 1 for details) once daily
and Group II was treated with an allopathy control formulation (see
Table 2 for details) once daily. All patients recruited were
screened and were determined to be suffering from stratum
granulosum in keratinization disorders. All patients were psoriasis
patients.
TABLE-US-00001 TABLE 1 Herbal Ointment Formula of Invention No.
Ingredient Quantity 1 Wrightia Tinctoria 5% 2 Cocos Nucifera 65% 3
Bees Wax 6% 4 Liquid Paraffin 24%
TABLE-US-00002 TABLE 2 Dithranol Ointment (Allopathy Control) No.
Ingredient Quantity 1 Dithranol 1% 2 Standard Ointment Base QS
[0042] Assignment of patients to treatment groups was randomized as
per standard statistical methods to minimize bias in the study.
Patients were enrolled in the study on a first come, first served
basis and assigned a subject number sequentially. The assignment of
each patient to the treatment group was determined by the
randomization list provided by the statistician.
[0043] Each patient voluntarily enrolled in the study and received
the treatment for 8 weeks. Skin biopsies at the treatment site were
taken from all patients at the beginning (T0) and end of the study
(T8w) for histopathological evaluation. In addition, at the
beginning (T0), end of treatment (T8w) haemogram analysis, liver
function testing and renal function testing were done to document
the safety profile of the treatments administered.
[0044] Histopathology of the skin biopsy was done by an expert
pathologist and the stratum granulosum parameter was measured at
visits T.sub.0 and T.sub.8w. The results of the stratum granulosum
measurements were scored as follows: [0045] (3)=representing the
absence of a granular layer [0046] (2)=representing a thinning of
the granular layer [0047] (1)=representing a normal thickness
granular layer of normal thickness. The stratum granulosum
parameter represents the thickness and integrity of the stratum
granulosum layer. The more active the disease, the thinner the
stratum granulosum layer and the lower the lipid content.
[0048] FIG. 2 presents photomicrographs of the skin of patients
observed before and after the 8-week treatment with the herbal
formulation of Wrightia tinctoria and Cocos nucifera. It is clear
from the photographs that the treatment with the herbal formulation
is very effective in increasing the integrity and thickness of the
stratum granulosum layer as compared to the condition prior to the
start of treatment.
Statistical Analysis of Results
[0049] Results of the statistical analysis of the stratum
granulosum measurement data for the two treatment groups are
presented in Table 3. A p-value of 0.05 is considered to be
significant.
TABLE-US-00003 TABLE 3 Statistical Analysis of Histopathology
Measurements for Stratum Granulosum Herbal Allopathy Control TIME
(Group I) (Group II) POINTS MEAN SD MEAN SD t P-Value T0W 2.00 0.94
1.6 0.84 1.000 0.331 T8W 1.00 0.00 1.2 0.63 1.000 0.331 PAIRED t
3.354 1.078 STATISTIC SIG 0.008 0.309 (2 - TAILED)
[0050] To examine the treatment effects, if any, a t-test was
performed with the data taken for the two groups at the beginning
and the end of the treatment. No statistical significance was
observed (p>0.05) for treatment effects on the stratum
granulosum measurements at the beginning (p=0.388) and the end of
treatment. That is, no difference in values could be attributed to
the different treatments).
[0051] To examine the time effects within each group, a paired
t-test was done with data at the beginning and end of treatment
within each group. With the herbal group, there was a statistically
significant time effect (p-values equal to 0.015) on the stratum
granulosum measurements and it was found that the stratum
granulosum values decreased with time suggesting a positive
response to herbal treatment with time.
[0052] However, with the allopathy control (Group II), no
statistically significant time effect was found for the Allopathy
control formulation (p-value equal to 0.081).
[0053] The statistical data analysis clearly indicates that the
herbal treatment for regression of stratum granulosum in
keratinization disorders is effective and is superior to the
allopathy control formulation.
Safety Evaluation
[0054] The safety of the use of the herbal formulation over the
treatment period was evaluated by taking measurements of vital
signs, haemogram measurements, liver function test (LFT)
measurements, and renal function test (RFT) measurements and
analyzing the data as a function of time.
[0055] The vital signs were measured 6 times during the treatment:
T0, T1w, T2w, T4w, T6w, and T8w; the haemogram, the LFT and RFT
measurements were made only at the beginning and end of the
treatment (T0, T8w).
[0056] The results of the statistical analysis of the vital sign
measurements (Systolic and Diastolic BP, pulse rate and respiratory
measurements) are presented in Table 4. The BP was measured using a
manual mercury sphygmomometer. The unit of measurement is mm of Hg.
The pulse rate was measured (beats per minute) in the radial artery
by palpating the artery with the middle, index and ring finger. The
respiratory rate was measured by watching the expansion of the
abdomen with each respiration and counting the expansions for one
minute.
TABLE-US-00004 TABLE 4 Statistical Analysis of Vital Sign
Measurements for herbal treatment. Respiratory Time BP-Systolic
BP-Diastolic Pulse Rate Rate Points Mean SD Mean SD Mean SD Mean SD
0w 121.10 15.31 81.00 8.76 87.60 17.33 23.00 6.20 T1w 111.40 11.43
77.00 8.01 75.80 8.77 21.40 7.00 T2w 114.00 14.30 79.20 9.85 74.60
11.70 22.30 6.93 T4w 107.00 8.23 79.00 5.68 85.40 11.47 24.20 5.45
T6w 111.40 8.00 78.80 5.27 78.70 22.60 24.00 3.62 T8w 109.00 12.87
78.00 6.32 82.40 11.96 25.40 4.99 Grand 112.32 12.36 78.83 7.28
80.75 14.88 23.38 5.72 Mean 1-Way 1.674 0.317 1.273 0.612 ANOVA
F-value p-value 0.157 0.901 0.289 0.691
[0057] A regular one-way ANOVA was also used to analyze the data at
different time points for the vital signs measurements. The data
clearly indicates that there were no statistically significant time
effects on BP systolic measurements (p=0.157); BP diastolic
measurements (p=0.901); pulse rate measurements (p=0.289) and
respiratory rate measurements (0.691) with the herbal treatment. In
summary, there is no statistically significant change in the vital
sign measurements over time due to treatment with the formulation
of the present herbal invention for the regression of stratum
granulosum in keratinization disorders, suggesting no safety
issues.
[0058] Results of the statistical analysis of the haemogram
measurements [Total count of white blood cells (TC), differential
white blood cells count as polymorphonuclear neutrophil (DC-P),
lymphocytes (DC-L), eosinophils (DC-E) and haemoglobin (Hb)] are
presented in Table 5. TC (Total count of white blood cells in the
blood) was measured using a Neubauer Counting Chamber. The normal
range for TC measurements is 4000-11,000 cells per cubic
millimeter. DC-P, which stands for the percentage of
P-polymorphonuclear neutrophil, was measured using Neubauer
Counting Chamber. The normal range for DC-P measurements is 55-65%
of total white cell count. DC-L, which is the percentage of
lymphocytes present, was measured using a Neubauer Counting
Chamber. The normal range for DC-L Measurements is 30-40% of the
total white cell count. DC-L was measured. DC-E, which is the
percentage of eosinophils, was measured using the Neubauer Counting
Chamber. The normal range for DC-E measurements is 1-7% of the
total white blood cell count. DC-E was measured. HB, which is the
haemoglobin measurements, was measured using the RA 50 Biochemical
Analyzer and the normal range is 12-14 gms.
TABLE-US-00005 TABLE 5 Statistical Analysis of Haemogram
Measurements for the herbal treatment. Time TC DC-P DC-L DC-E HB
Points Mean SD Mean SD Mean SD Mean SD Mean SD T0w 7343.00 1588.76
57.30 2.95 37.90 1.79 4.80 2.90 13.02 1.72 T8w 8634.00 1104.94
58.90 2.69 37.10 2.38 4.00 2.49 12.95 0.94 Paired in -1291.00
2279.49 -1.60 3.78 0.80 3.22 0.80 3.77 0.075 2.13 differ mean
Paired t -1.791 -1.340 0.784 0.672 0.100 statistic Sig 0.107 0.213
0.453 0.519 0.924 (2-tailed)
[0059] To examine the time effects, a paired t-test was done with
data taken at the beginning and end of treatment for each of the
haemogram measurements. The data clearly indicates that there were
no statistically significant time effects on TC measurements
(p=0.107); DC-P measurements (p=0.213); DC-L measurements
(p=0.453); DC-E measurements (p=0.519) and HB measurements
(p=0.924) with the herbal treatment. In summary, there is no
statistically significant change in haemogram measurements with
time due to the treatment with the herbal formulation, suggesting
no safety issues.
[0060] Results of the statistical analysis of the liver function
test (LFT) measurements [serum glutamic oxalo acetic transaminase
(SGOT), serum glutamic pyruvic transaminase (SGPT) and serum
bilirubin] are presented in Table 6. SGOT, serum glutamic-oxalo
acetic transaminase (international unit per liter), was measured at
visits T.sub.0 and T.sub.8w. The normal range is 0-46 IU/L. SGPT,
serum glutamic pyruvic transaminase (international units/liter) was
measured at visits T.sub.0 and T.sub.8w. The normal SGPT ranges
from 0 to 49 IU/L. The serum bilirubin was measured at visits
T.sub.0 and T.sub.8w. The normal serum bilirubin ranges from 0.0 to
1.0 mg/dl.
TABLE-US-00006 TABLE 6 Statistical Analysis of Liver Function Test
(LFT) Measurements for the herbal treatment. Time SGOT SGPT Serum
Bilirubin Points Mean SD Mean SD Mean SD T0w 24.90 8.80 26.10 14.78
0.73 0.23 T8w 24.00 8.94 26.60 11.01 0.69 0.24 Paired in 0.90 10.97
-0.50 11.24 0.035 0.31 differ mean Paired t 0.259 0.141 0.352
statistic Sig 0.801 0.891 0.733 (2-tailed)
[0061] To examine the time effects a paired t-test was done with
data taken at the beginning and the end of treatment for each of
the LFT measurements. The data clearly indicates that there were no
statistically significant time effects on SGOT measurements
(p=0.801); SGPT measurements (p=0.891); and the serum bilirubin
measurements (p=0.733) with the herbal treatment. In summary, there
is no statistically significant change in LFT measurements with
time due to treatment with the herbal formulation of the present
invention for the regression of stratum granulosum in
keratinization disorders, suggesting no safety issues.
[0062] Results of the statistical analysis of the Renal Function
Test (RFT) measurements for serum creatinine and serum urea,] are
presented in Table 7. Serum creatinine was measured at visits
T.sub.0 and T.sub.8w. The normal serum creatinine value ranges from
0.8 to 1.4 mg/dl. Serum urea was measured at visits T.sub.0 and
T.sub.8w. The normal serum urea value ranges from 10 to 50
mg/dl.
TABLE-US-00007 TABLE 7 Statistical Analysis of Renal Function Test
(RFT) Measurements for the Herbal Treatment. Time Serum Creatinine
Serum Urea Points Mean SD Mean SD T0w 1.06 0.22 32.40 17.50 T8w
1.08 0.18 25.49 7.75 Paired in -0.021 0.244 6.91 18.81 differ mean
Paired t -0.271 1.161 statistic Sig 0.792 0.275 (2-tailed)
[0063] To examine the time effects paired t-test was done with data
at the beginning and end of treatment for each of the RFT
measurements. The data clearly indicates that there were no
statistically significant time effects on serum creatinine
measurements (p=0.792) and serum urea measurements (p=0.275) with
the herbal treatment. In summary, there is no statistically
significant change in RFT measurements with time due to treatment
with the herbal formulation of the present invention for the
regression of stratum granulosum in keratinization disorders,
suggesting no safety issues.
[0064] It is clear from the histopathological examination and
statistical analysis of the clinical data that the herbal
formulations of the compositions of the present invention are very
effective in the treatment of stratum granulosum and is superior to
the allopathy control. In addition, the evaluation of haemogram,
LFT and RFT test results clearly show that the herbal formula of
the present invention is also safe to use on humans when used as
directed.
[0065] Other modifications and variations of the present invention
will become apparent to those skilled in the art from an
examination of the above specification and examples. Therefore,
other variations of the present invention may be made which fall
within the scope of the appended claims even though such variations
were not specifically discussed above.
* * * * *