FORM 2
THE PATENTS ACT, 1970
THE PROVISIONAL SPECIFICATION
(See section 10)
1. THE PROCESS OF PREPARING THE TOPICAL ANTI-INFLAMMATORY ANALGESIC PREPARATION.
2. Cadila Pharmaceuticals Limited, IRM House, Off C.G. Road, Navrangpura, Ahmedabad-380009, Gujarat, India, an Indian company.
3. The following specification particularly describes and ascertains the nature of this invention

THE PROCESS OF PREPARING THE TOPICAL ANTI-INFLAMMATORY /ANALGESIC
PREPARATION.
Inflammation and subsequent pain in various parts of the body can lead to temperory or permanent disability and/or difficulty in carrying out day to day activity. This can be treated by providing drugs through systemic route or by topical application of medicines. Advantages of topical route include:
1. ease of administration of drugs which cannot be given systemically.
2. capability to achieve higher local concentration and lower systemic concentration, thus improving therapeutic ratio of drug and decreasing side effect profile.
The purpose of the present invention is to provide a process of preparing the topical formulation which relieves inflammation and pain effectively at local site. The further objective of present invention is to provide a process of preparing the topical formulation which is non-irritating.
According to present invention, a topical preparation can be prepared using Guggulu, Nirgundi, Asthisamhara, Bramhatirtha, Nimba, Amalakee, Haritaki, Bibhitaka, Karpura, Sarala, Ajmoda extract, Devdaru tel, Nilgiri tel and Cajuputi tel and other ingredients. The above ingredients may be combined with known anti-inflammatory agents like diclofenac, nimesulide etc.
The preparation made according to present invention has been evaluated in adjuvant -induced arthritis in rats. It is found to significantly inhibit edema in comparison to control group. Inhibition was found to be approximately 60% at day 4 (p < 0.05) and 66.87% at day 7 (p < 0.001).
In carrageenin edema model, the preparation made according to present invention was found to inhibit edema by 90.10%.

In humans suffering from osteoarthritis, the preparation made according to present invention resulted in decrease in pain at rest, use of joints. There was found to be improvement in range of movement also.
Thus, topical preparation made according to present invention can be useful in the management of inflammatory conditions of various parts of the body by topical application. It can also be used when inflammation/pain is secondary to degenerative conditions like osteoarthritis.
Dated this 23rd April, 2001 Dr. Bakulesh M Khamar

THE PROCESS OF PREPARING THE TOPICAL ANTI-INFLAMMATORY AND ANALGESIC PREPARATION
This invention relates to the process of preparing the topical anti-inflammatory and analgesic preparation.
BACKGROUND AND PRIOR ART RELATING TO THE INVENTION:
Plants and herbs have long been used for medicinal purposes. Indeed, India has been known since long of the healing powers of certain herbs as remedies for various illnesses. The present invention provides the process of preparing the topical anti-inflammatory preparation for the use in the treatment of inflammatory conditions of rheumatoid arthritis, osteoarthritis, cervical and ankylosing spondylitis, and painful conditions of frozen shoulder, stiff neck, back aches and musculoskeletal disorders.
Typically, herbal medicines are obtained as the active compound(s) by extraction process from plant tissues. Natural ingredients, e.g., herbs, have a|so been used to treat bone and joint inflammation, especially in India, and, increasingly, in western countries.
The Ayurvedic system of Indian traditional medicine provides many plants for the treatment of inflammatory disorders in human beings and animals. Majority of these plants have been investigated individually for their beneficial medicinal properties.
Commiphora mukul (Guggul): Commiphora mukul (C.mukul) is grown in the arid and rocky zones in certain parts of Southwest and Northwestern region of India. The Oleoresin from commiphora mukul contains 0.37% essential oil containing mainly myrecene, dimyrecene, and polymyrecene. The steroidal component of petroleum ether extract of c. mukul has marked antiarthritic effect, comparable to that of hydrocortisone and more potent than phenylbutazone (1). It has a high anti¬inflammatory potential against Brownlee's formaldehyde induced arthritis in albino rats (2). Carragenan oedema of the rat paw also regressed on exposure to the steroidal component of petroleum ether extract of C.mukul (3,4). The gum resin from C.mukul has been used in the treatment of various clinical types of arthritis (5). The aqueous extract of C.mukul significantly inhibited maximum and total oedema response of carragenan-induced rat paw edema (6).
Vitex negundo (Nirgundi) :. Vitex negundo is grown commonly in Wastelands, on the roadside, on the banks of streams or in moist places near decidous forests. The ethylacetate extract of leaves showed a significant activity against carragenan. 5-HT and bradykinin-induced inflammatory oedema. It also possessed significant inhibitory effect against granuloma pouch and cotton pellet implantation though less potent than phenylbutazone and beta-methasone(7,8). Patients with rheumatoid arthritis showed encouraging results when treated with decoction of nirgundi plant(9).
Cissus quadrangularis (Asthisamhara): The plant cissus quadrangularis is a creeper common in the hotter parts of India, used in folklore medicine to heal bone fractures. The effect of the plant on the healing of fractures has been extensively studied by Udupa and Prasad (10-13). The anabolic phytosteroid has been shown to markedly

induce an early regeneration of the connective tissue and rapid mineralisation of callus. The extracts when administered either intra-muscularly or locally, reduces the healing time of fractures (14).
Costus speciosus (Bramhatirtha): The plant costus speciosus is distributed more or. less throughout India. The root is pungent, bitter useful in inflammation and rheumatism (15).
Melia azadirachta (Nimba) : Melia azadirachta is a indigenous tree grown in all the plains in the Indian subcontinent. Seed oil and Kernel oil are useful in rheumatism and sprains when applied locally (16).
Emblica Officinalis (Amalakee) : Emblica officinalis grows throughout India upto 4500 ft2. The water fraction, of methanolic extracts of leaves of Emblica officinalis significantly inhibited the edema formation induced by carrageenin and dextran in rat hind paw. The anti-inflammatory mechanism of this extract is through its inhibitory action against the migration of polymorphonuclear leucocytes (17).
Terminalia chebula (Haritaki) : Terminalia chebula grows below an elevation of 1000 m above sea level throughout India except in dry and arid regions of Rajasthan.
Terminalia Belerica (Bibhitaka): Terminalia Belerica grows throughout the forests of India, Burma, Srilanka except in the dried and arid regions.
Emblica officinalis, Terminalia chebula and Terminalia Belerica prevent aging and impart longevity, immunity, and enhance body resistance against disease (18).
Camphora officinarum (Karpura): Comphora officinarum is commonly distributed throughout India, Japan and China. It can be used externally in the cases of inflammations, bruises and sprains.
Pinus longifolia (Sarala) : Pinus longifolia is found in Himalayas at altitude of 450-2400 m. It grows well in the plains. It can be used as a rubificient in rheumatism and for deep-seated inflammation.
The purpose of the present invention is to provide a process of preparing the topical formulation, which relieves inflammation and pain effectively at local site without producing any local irritation.
Several herbal topical preparations are available in India for the treatment of pain and inflammation, which are described as follows:
Rheuma oil from Baidyanath consisting decoction derived from Arun mookl, Dhatura panchang, Ashwagandha, Satavari, Amala, Haldi, Kuchala, Vastanabh prasarini each 3% w/v, kapur 5% w/v. Tarpio oil 10% w/v Gandh purna oil 25% v/v in Til oil base.
Rhumasy in the form of oil and ointment from Zandu consisting a preparation from Narayan oil 2.5 ml, Vishagarbha oil 2.5 ml, Maha Mash oil 2.5 ml, and Gandhpuro oil 2.5 ml.

Rumalaya cream from Himalaya drug Co. consisting of oil of Karpoora 5%, Putiha 10%, Sarala 5%, Thwak 2.5%, Oil processed with Jatamansi, Kushtha, Jyotishmati, Nirgundi, Vishnu priya, Vacha, Yavani, Akarakarabha, Alarka, Dhatura, Rasna, Laksha, Maricha, Devdaru, Daruharidra, Gandhaava hasta, Manjistha, Chitraka, Karavira, Dashamoola 22% in a cream base.
Rheumatil oil and gel from Dabar consisting of Mahanarayan oil 70%, Gandhapuro oil 10%, Pudina satva 3%, Neelgiri oil 2%, Lavang oil 2%, Guggula satva 2%, Sunthi satva 1% in a oil or gel base.
Compositions composed of. natural ingredients and said to be useful in reducing inflammation are disclosed in several US patents, e.g., in U.S. Pat. Nos.5,120,538, 5,494,668, 5,683,698, 5,707,631, 5,916,565, 5,888,514, 5,908,628; 5,788,971; 5,854,291; and 5,910,307, and in Japanese Patent No. 4,005,237 as well as in German patent No. 3,724,341.
U.S. Pat. No. 5,120,538 is directed to a method of treating inflammation in a patient by administration of an effective dose of a pharmaceutical composition comprising essential oils extracted from tissues of Curcuma domestica, or Curcuma xanthorrhiza or both oils and curcuminoid substantially free of bis-desmethoxycurcumin.
U.S. Pat. No. 5,707,631 is directed to a therapeutic herbal composition including Trigonella foenum-graecum seed, Syzygium aromaticum fruit, Allilum sativum bulb, Cinnamomum zeylanicum bark, Saussurea costus root and Euphorbia lathyris bud together with sodium chloride (preferably sea salt).
U.S. Pat. No. 5,494,668 (Patwardhan) discloses a method of treating degenerative musculoskeletal diseases such as rheumatoid arthritis and osteoarthritis in an animal, typically a human, involving administering to the animal, typically enterally, in a convenient dosage form, a therapeutically effective amount of the beneficiated extracts of the plants Ashwagandha, Sallai Guggul,.. Turmeric, and Ginger, in a predetermined propprtion relative to each other with or without other biologically active inorganic ingredients.
U.S. Pat. No. 5,683,698 (Chavali et al.) discloses an herbal formulation and its use for reducing/alleviating symptoms associated with rheumatoid arthritis, osteoarthritis, and reactive arthritis and for reducing the production of pro-inflammatory cytokines. The formulation contains an herbal extract from the roots, rhizomes and/or vegetation of six herbal plant varieties, specifically, the species of Alpinia, Smilax, Tinospora, Tribulus, Withania, and Zingiber.'The patent further discloses foods, beverages and medicaments in the form of capsules, tablets, liquids, and the like, containing the herbal formulation.
U.S. Pat. No. 5,916,565 (Rose et al.) discloses an orally administered composition for prophylaxis and therapy of joint and connective tissue disorders in vertebrates, wherein the composition contains metabolic precursors, herbal phytochemicals, and palatability agents. Suitable herbal phytochemicals are said to include cayenne, ginger, turmeric, yucca, Devil's claw, nettle leaf, Black Cohosh, alfalfa and celery seeds.

U.S. Pat. No. 5,888,514 (Weisman) discloses a composition for treating bone or joint inflammation in mammals, wherein the composition contains a systemically absorbable cartilage and an amninosaccharide and may optionally contain, among other ingredients, one or more extracts of an herb of the genus Withenia, of the bark of an herb of the genus Salix, or of a root of an herb of the genus Panax.
U.S. Pat. No. 5,908,628 (Hou) discloses compositions for treating osteoarthritis and rheumatoid arthritis, containing talc, silkworm excrement, and ingredients of plants of species of the genera Stephania, Coix, Pinellia, Prunus, Phellodendron, Sophora, Tetrapanax, Stemona, Glycyrrhiza, Tripterygium, Forsythia, and Siegesbeckia.
U.S. Pat. No. 5,788,971 (Togasaki) discloses an active oxygen free radical scavenging agent composed of green tea leaf extract containing epigallo catechin gallate and sunflower seed extract containing chlorogenic acid.
U.S. Pat. No. 5,854,291 (Laughlin, et al.) discloses a topically-applied pain reliever composition for treating such discomforts as arthritis pain, composed of capsaicin and, optionally, a plant extract selected from the group consisting of nettle extract, yarrow extract, coltsfoot extract, birch extract, rosemary extract, horsetail extract, ginger extract, chamomile extract, comfrey extract, lavender extract, and bergamot extract.
U.S. Pat. No. 5,910,307 (Kwak, et al.) discloses a combined medicinal plant composition for alleviating acute/chronic inflammation, composed of Clematis Radix, Trichosanthes root, and Prunella Herba (which contains oleanolic acid ursolic acid) in a certain ratio.
Japanese Patent Publication No. 4,005,237 discloses a combination of Cinnamomum sieboldii and Allium sativum for superoxide scavenging in the treatment of inflammatory disorders. German patent Publication No. 3,724,341 discloses a combination of Cinnamomum zeylanicum, Pumica granitum cortex, Cardamon zingiberaceae fruit and Piper longum fruit.
Although the use of various herbs have been described in related areas, the present invention provides a process of preparing the topical anti-inflammatory and analgesic preparation which comprises a different group of herbs with synergistic activity if •used as per the present invention.

REFERENCES:
1. Sharma JN and Jain JC. Indian J Pharmacol, 1978, 10:87.
2. Gujral ML et al. Ind J Physiol Pharmacol I960, 4: 267-273.
3. Arora RB et al. Ind J Exp Biol 1971, 9: 403.
4. Arora RB et al. Ind J Med Res 1972, 60: 920.
5. Satyavati GV et al. Rheumatism 1969, 4: 141.
6. DuV/iejua M et al. Planta Medica 1993, 59: 12-16.
7. Sharma AK and Singh RH. Bull Med Ethnobot Res 1980, 1: 262.
8. Mohiddin SG. Rheumatism 1974, 14: 97.
9. Jain PK and Pande TK. J Res Ind Med Yoga Homeo 1976, 11:2.
10. Prasad GC and Udupa KN. J Res Ind Med 1972, 7(4): 29-34.
11. Chopra SS et al. Ind J Med Res 1975, 63: 824-828.
12. Udupa KN and Prasad GC. Ind J Med Res 1964, 52: 26.
13. Prasad GC and Udupa KN. Ind J Med Res 1963, 51: 667-676.
14. Udupa KN and Prasad GC. Ind J Med Res 1964, 52: 480-487.
15. Kirtikar KR, Basu BD. Indian Medicinal Plant Vol.IV p.2440-41.
16. The treatise on Indian Medicinal Plants. Ed. Chatterjee A and Pakrashi SC. Vol.3. National Institute of Science Communication, New Delhi, 1997, p.76-79.
17. Asmawi MZ et al J Pharm Pharmacol 1993,45: 581-4.
Nadkarani KM. The Indian Materia Medica 3rd Ed. Vol.1, Popular Prakashan, Bombay, 1976.

OBJECTIVES OF THE PRESENT INVENTION:
It is an objective of the present invention to provide a process for preparing the topical anti-inflammatory and analgesic compositions based on the extraction process of herbal ingredients that has potential analgesic and anti-inflammatory activity such as frozen shoulder, stiff neck, backaches, and musculoskeletal pains and rheumatoid arthritis, osteoarthritis, cervical and ankylosing spondylitis and sciatica.
It is another objective of the invention to produce pharmaceutical compositions based on the process described herein, from the cissus quadrangularis stem powder and costus speciosus stem powder.
A further objective of this invention is to provide synergistic compositions based on the extraction process of herbal ingredients.
A further objective of this invention is to provide compositions aforesaid that act without exerting toxic or side effects.
DETAILED DESCRIPTION OF THE INVENTION:
The current invention in this application is the process for the preparation of compositions comprising of herbal ingredients.
The process for the preparation of topical anti-inflammatory and analgesic preparation essentially comprises of powders of cissus quadrangularis stem and costus speciosus stem. The process also comprises of powders of seedless fruit of terminalia chebula, terminalia bellarica, Embilica officinalis, melia azadirchta leaf, vitex negundo leaf, gum resin of commiphbora mukul. The process also optionally includes volatile oils like opium graveolens extract, pinus longifolia, camphora officinarum, cedrus deodara, Eucalyptus globules, and Melaleuca leucadendron.
The process of preparing the topical anti-inflammatory and analgesic preparation as per the present invention includes the following steps:
a) Extraction of herbal ingredients water;
b) Concentrating the extraction by boiling it;
c) Filtration of the concentrated solution to remove residue;
d) Addition of oil base to the filtrate;
e) Removal of solvent by further boiling the mixture of base oil and filtrate;
f) Resultant extract is allowed to cool;
g) Addition of volatile oils to solution.
The compositions derived from a process of present invention have synergistic action and are used to relieve all painful and inflammatory conditions like rheumatoid arthritis, osteoarthritis, cervical and ankylosing spondylitis and sciatica, frozen shoulder, stiff neck, bach aches and musculoskeletal pains.
The following examples describe the invention however; they should not limit the scope of invention.

Example I
Examples of types and amount of herbal ingredients in 60 ml used in preparation of topical anti-inflammatory / analgesic composition are described as below.

Ingredient
Extract of Cissus quadrangularis (Asthisamhara) Extract of Costus speciosus (Bramhatirtha),

Weight of ingredient In the composition 2.084 gm 5.289 gm

II
Ingredient
Extract of Cissus quadrangularis (Asthisamhara) Extract of Costus speciosus (Bramhatirtha)

Weight of ingredient In the composition 0.521 gm 4.171 gm

III
Ingredient
Extract of Cissus quadrangularis (Asthisamhara) Extract of Costus speciosus (Bramhatirtha)

Weight of ingredient In the composition 2.084 gm 1.763 gm

IV
Ingredient
Extract of Cissus quadrangularis (Asthisamhara) Extract of Costus speciosus (Bramhatirtha) Gum resin of Commiphora mukul(Guggulu)

Weight of ingredient In the composition 2.084 gm 5.289 gm 2.607 gm

V , Ingredient
Extract of Cissus quadrangularis (Asthisamhara) Extract of Costus speciosus (Bramhatirtha) Gum resin of Commiphora mukul(Guggulu)

Weight of ingredient In the composition 0.521 gm 4.171 gm 2.607 gm

VI
Ingredient
Extract of Cissus quadrangularis (Asthisamhara) Extract of Costus speciosus (Bramhatirtha) Gum resin of Commiphora mukul(Guggulu) Extract of Vitex negundo(Nirgundi)

Weight of ingredient In the composition 2.084 gm 5.289 gm 2.607 gm 6.257 gm

VII ,
Ingredient Weight of ingredient
In the composition
Extract of Cissus quadrangularis (Asthisamhara) 0.521 gm
Extract of Costus speciosus (Bramhatirtha) 4.171 gm
Extract of Melia azadirchta(Nimba) 2.607 gm
Extract of Vitex negundo(Nirgundi) 6.257 gm
.
VIII
Ingredient Weight of ingredient
In the composition
Extract of Cissus quadrangularis (A'sthisamhara). . 0.521. gm
Extract of Costus speciosus (Bramhatirtha) . 4.171 gm
Extract of Melia azadirchta(Nimba) . .607 gm
Extract of Vitex negundo(Nirgundi) 6.257 gm
Extract of Emblica officinalis(Amalaki) 1.0428 gm
Gum resin of Commiphora mukul(Guggulu) 2.607 gm
Extract of Terminalia Chebula(Haritaki) 1.0428 gm
Extract of Terminal] bellerica(Bibhitaka) 1.0428 gm
Ingredient Weight of ingredient
In the composition
. Extract of Terminalia Chebula(Haritaki) 1.0428 gm
' Extract of Terminali bellerica(Bibhitaka) 1.0428 gm
Extract of Emblica officinalis(Amalaki) 1.0428 gm
Extract of Melia azadirchta(Nimba) 2.607 gm
Extract of Vitex negundo(Nirgundi) 6.257 gm
Extract of Cissus quadrangularis(Asfhisamhara) 0.521 gm
Extract of Commiphora mukul(Guggulu) 2.607 gm
Extract of Costus speciosus(Bramhatirtha) 4.171 gm
X
Ingredient Percentage of ingredient
In the composition
Extract of Costus speciosus(Bramhatirtha) 11.83%
Extract of Melia azadirchta(Nimba) 6.3%
Extract of Terminali bellerica(Bibhitaka) 2.52%
Extract of Emblica officihalis(Amalaki) 2.52%
Extract of Commiphora mukul(Guggulu) 7.41 %
Extract of Vitex negundo(Nirgundi) 15.1 %
Extract of Cissus quadrangularis(Asthisamhara) 1.25%
Extract of Terminalia Chebula(Haritaki) 2.52%

Example II
Examples of types and amount of base oils used in preparation of topical anti¬inflammatory / analgesic compositions are describedas as below.

1. 2. 3. 4. 5. 6. 7. 8. 9.

Cotton seed oil
Sesame oil
Mustard oil
Cotton seed oil and Sesame.oil in a ratio of 2:3 Cotton seed oil and Mustard oil in a ratio of 7:3 Sesame oil and Mustard oil in a ratio of 7:3 Cotton seed oil and Sesame oil in a ratio of 1:4 Sesame oil and Mustard oil in a ratio of 3.5:6.5 Cotton seed oil and Mustard oil in a ratio of 1:3

Example HI
Examples of types and amount of volatile oils used in preparation of topical anti¬inflammatory /,analgesic composition are described as below.

I
Ingredient
Pinus longifplia (Sarala) Camphor (karpur) Apium graveolens (Ajmoda) Cedrus deodara (Devdaru oil) Eucalyptus globulus (Nilgiri oil) Melaleuca leucadendron

Weight of ingredient In the composition 2.600 gm 3.259 gm 2.600 gm 1.629 gm 1.629 gm 1.629 gm

II
Ingredient
Pinus longifolia(Sarala)
Camphor(karpur)
Apium graveolens(Ajmoda)
Cedrus deodara(Devdaru oil)
Eucalyptus globulus (Nilgiri oil)
Melaleuca leucadendron

Weight of ingredient In the composition 1.354 gm 3.259 gm 4.062 gm 0.529 gm 1.629 gm 2.644 gm

III
Ingredient
Pinus longifolia(Sarala)
Camphor(karpur)
Apium graveolens(Ajmoda)
Cedrus deodara(Devdaru oil)
Eucalyptus globulus (Nilgiri oil)
Melaleuca leucadendron

Weight of ingredient In the composition 4.062 gm 1.058 gm 2.354 gm 2.644 gm 0.529 gm 1.529 gm

IV
Ingredient
Pinus longifolia(Sarala) Apium graveolens(Ajmoda) Cedrus deodara(Devdaru oil) Eucalyptus globulus (Nilgiri oil)
V Ingredient
Camphor(karpur) Apium graveolens(Ajmoda) Cedrus deodara(Devdaru oil) Eucalyptus globulus (Nilgiri oil) Melaleuca leucadendron
VI
Ingredient
Apium graveolens(Ajmoda) Cedrus deodara(Devdaru oil) Eucalyptus globulus (Nilgiri oil)

Weight of ingredient In the composition 2.600.gm 2.600 gm
1.629 gm 1.629 gm
Weight of ingredient
In the composition .
3.259gm 2.600gm 1.629 gm 1.629 gm 1.629 gm
Weight of ingredient In the composition 2.600 gm 1.629 gm 1.629 gm

Example IV
Examples of types of processes describing Extraction, Concentrating the extract, Filtration, Addition of oils and cooling procedures used in preparation of topical anti¬inflammatory / analgesic composition are described as below.
Process I:
Mix herbs Terminalia chebula, Terminalia bellarica, Emblica officinalis, Melia azardichta, Vitex negundo, Cissus quadrangularis, commiphora mukul, and costus speciosus. Add solvent, which weighs 20 times as much as the total weight of herbs into a mixture of herbs. Boil the mixture at 80 °C till the volume of solvent reduced to approximately half. After boiling filter the extract to get clear liquid. This filtrate was mixed with 3 times weight of base oil by weight of herbal ingredients and boil at 80 °C till all solvent gets removed and oil becomes completely free from solvent. The oily solution was allowed to cool. In a cool oily solution add pinus longifolia,, camphora officinarum, Apium graveolens, Cedrus deodara, Eucalyptus globulus and Melaleuca leucadendron. Mix thoroughly to get oily liquid.
Process II:
Mix herbs Terminalia chebula, Terminalia bellarica, Emblica officinalis, Melia azardichta, Vitex negundo, Cissus quadrangularis, commiphora mukul and costus

speciosus. Add solvent, which weighs 20 times as much as the total weight of herbs into a mixture of herbs. Add 3 times weight of base oil by weight of herbal ingredients. Boil the mixture at 80 °C till volume of solvent reduced to half. After boiling filter the extract to get clear liquid. This filtrate was further boiled at 80 °C till all solvent gets removed and oily solution becomes completely free from the solvent. The oily solution was allowed to cool. In a cool oily solution add pinus longifolia, camphora officinarum, Apium graveolens, Cedrus deodara, Eucalyptus globulus and Melaleuca leucadendron. Mix thoroughly to get, oily'liquid'.
Process III:
Mix herbs Terminalia chebula, Terminalia bellarica, Emblica' officinalis, Melia azardichta, Vitex negundo, Cissus quadrangularis, and commiphora mukul. Add solvent, which weighs 20 times as much as the totafweight of herbs into a mixture of herbs. Boil the mixture at 80 °C till volume of solvent reduced to half. After boiling filter the extract to get clear liquid. This filtrate was mixed with 3 times weight of base oil by weight of herbal ingredients and boil at 80 °C till all solvent gets removed and oil becomes completely free from solvent. This is designated as Extract I. Take the herb costus speciosus and repeat the process as described for Extract I. The resultant oily solution is designated as Extract II.
Mix the Extract I and Extract II and allow them to cool. In a cool oily solution add pinus longifolia, camphora officinarum, Apium graveolens, Cedrus deodara, Eucalyptus globulus and Melaleuca leucadendron. Mix thoroughly to get oily liquid.
Example V:
Examples of physicochemical characteristics of pharmaceutical compositions:
The herbal compositions derived from the process of present invention are also evaluated for its stability. The herbal compositions are evaluated at different test conditions of temperature and humidity (25° C / 60% RH for 36 months, and 40° C / 75% RH for 6 months). It was found that the herbal compositions are stable at room temperature as well as at accelerated condition for physicochemical characteristics and therapeutic efficacy.
NON-HUMAN ANIMAL TRIALS:
Pharmacologic research shows that the process for the preparation of compositions of the present invention has an anti-inflammatory effect on arthritis caused by Freund's adjuvant in rats. The compositions of the present invention also provide anti¬inflammatory activity due to carrageenin-induced inflammation in rats.
Method of Evaluation:
I. Fraund's Adjuvant induced Arthritis in Rats:
Arthritis was induced in eight groups of 6 rats (Female Wistar, 180-200g, 8 week old), each by an intradermal injection of 0.05 ml of complete Freund's adjuvant (Sigma Laboratory, USA) in the right hind footpad. Six rats were not injected and served as nonadjuvant controls. All the seven preparations were locally applied

once daily to the injected footpad for 7 days. The inhibitory effect of test
compositions was assessed by measurement of the paw volume by the volume
displacement method (Plethysmometer, UGO BASILE, Italy) immediately before
injection of complete Freund's adjuvant and at 4th and 7th day after the application.
A group of six rats were not applied any preparation and served as positive
control.

*, - /
II. Carrageenin induced. Acute Inflammation in Rats:
Acute inflammation was induced in eight groups of 4 rats (female Wistar rats, 180-200g, 8-9 weeks old), each by injecting 0.1 ml of carrageenin (1% w/v) solution in normal saline beneath the sub plantar region of the right hind paw. The paw volume of rats was measured with a plethysmometer (model 7140, UGO BASAILE, Italy) immediately before and at 2nd, 3rd and 4th hour after carrageenin injection. All the seven compositions were locally applied half an hour after carrageenin application. The mean increases in paw volume were calculated and results are shown as percentage inhibition of swelling compared with the control.
I. Fraund's Adjuvant induced Arthritis in Rats: The following examples
1-4 describe the evaluation of various herbal formulations in the treatment of arthritis caused by Fraund's adjuvant in rats.
Example 1:
Evaluation of the composition derived from the process of the present invention comprises of extract of Cissus Quadrangularis & Costus Speciosus in base oil. This composition now designated as composition 1, which has been evaluated in the treatment of arthritis caused by Freund's adjuvant in rats.
Single injection of 0.05 ml complete Freund's adjuvant produced a significant ' increase in paw volume as compared to nonadjuvant control at day 4 that is also maintained on day 7.
The protective effect, as calculated by percentage decrease in the paw volume, was found to be significant with composition 1 as compared to adjuvant control. Composition 1 has been found to decrease in paw volume by 40% and 36% on day 4 and day 7 respectively after single application in a day up to seven days. The results demonstrating the efficacy of herbal compositions 1 on the protection of arthritis have been shown in Table: 1.
Example 2:
Evaluation of the composition derived from the process of the present invention comprises of extract of Cissus Quadrangularis, Costus Speciosus, Terminalia chebula, Terminalia bellarica, Emblica officinalis, Melia azardichta, Vitex negundo, and commiphora mukul'in base oil along with volatile oils pinus longifolia, camphora officinarum, Apium graveolens, Cedrus deodara, Eucalyptus globulus and Melaleuca leucadendron. This composition now designated as composition 2, which has been evaluated in the treatment of arthritis caused by Freund's adjuvant in rats.

Single injection of 0.05 ml complete Freund's adjuvant produced a significant increase in paw volume as compared to nonadjuvant control at day 4 that is also maintained on day 7.
The protective effect, as calculated by percentage decrease in the paw volume, was
found to be significant with composition 2 as compared to adjuvant control.
Composition 2 has been found to decrease in paw volume by 36% and 42% on day 4
and day 7 respectively after single application in a day up to seven days. The results
demonstrating the efficacy of herbal composition 2 on the protection of arthritis have
been shown in Table: 1.

Table 1

Model : Fraund's Adjuvant
induced arthritis 0.05 ml in right
paw
% decrease in Paw volume (right paw) as compared to control
Group Day 4 Day7
Composition 1 40.39 36.12
Composition 2 34.36 42.70
Example 3:
Evaluation of the composition derived from the process of the present invention comprises of extract of Cissus Quadrangularis or Costus Speciosus in base oil- This composition, now designated as composition 3 and composition 4, which has been evaluated in the treatment of arthritis caused by Freund's adjuvant in rats.
The compositions containing only Cissus Quadrangularis or Costus Speciosus (Composition 3 and 4) produced a 20% and 30% decrease in paw volume as compared to adjuvant control in rats with adjuvant-induced arthritis. This effect is less than the composition 1 which contains both Cissus Quadrangularis and Costus Speciosus. These results indicate that the costus -and cissus when combined together give better effect. The results demonstrating the efficacy of herbal compositions 3 and 4 on the protection of arthritis have been shown in Table: 2.
Table : 2

Model: Fraund's Adjuvant
induced arthritis 0.05 ml in right
paw
% decrease in Paw volume (right paw) as compared to control
Group Day 4 Day7
Composition 3 29.72 33.84
Composition 4 23.99 19.23

Example 4
Removal of Cissus Quadrangularis and Costus Speciosus both or Cissus
Quadrangularis or Costus Speciosus from the process for preparing composition 2 has
also been evaluted.. These composition 7 respectively, which have been evaluted in the
composition 6 and composition 7 respectively, which have been evaluated in the
treatment of arthritis caused by Freund's adjuvant in rats.' ,
The removal of cissus or costus, as in composition 6 and 7, from composition 2 described above leads to a drastic reduction in the activity, however, removals of both the ingredients provide a significant decrease in. a paw volume on day 7 as compared to adjuvant control. Composition 6 and 7 infact, does not produced any activity. However composition 5 decreased the paw volume by 26% on day 7 that is significantly different as compared to adjuvant control. These results also confirm the essentiality of Costus and Cissus in the composition and also the synergistic action of both the ingredients. The results demonstrating the effect of herbal compositions 5, 6 and 7 on the protection of arthritis have been shown in Table: 3.
Table : 3

Model : Fraund's Adjuvant
induced arthritis 0.05 ml in right
paw
% decrease in Paw volume (right paw) as compared to control
Group Day 4 Day7
Composition 5 12.32 26.09
Composition 6 6.64 1.26
Composition 7 2.99 0.77
II. Carrageenin induced Acute Inflammation in Rats: The following examples 1-4 describe the evaluation of various herbal formulations in the treatment of acute inflammation caused by carrageenin in rats.
Example: 1
Evaluation of the composition derived from the process of the present invention comprises of extracts of Cissus Quadrangularis & Costus Speciosus in base oil. This composition now designated as composition 1, which has been evaluated in the treatment of acute inflammation caused by carrageenin in rats.
Single injection of 0.1 ml of Carrageenin produced a significant increase in paw volume as compared to control after 2nd , 3rd and 4th hours.
The protective effect, as calculated by percentage decrease in the paw volume, was found to be significant with composition 1 as compared to carrageenin control. Composition 1 has been found to decrease in paw volume by 50% at 2nd and 3rd hours and 36% at 4th hour respectively after single application. The results demonstrating

the efficacy of herbal composition shown in Table: 4.

on the protection of inflammation has been

Example 2:
Evaluation of the composition derived from the process of the present invention comprises of extract of Cissus Quadrangularis, Costus Speciosus, Terminalia chebula, Terminalia bellarica, Emblica officinalis, Melia azardichta, Vitex negundo, and commiphora mukul in base oil along with volatile oils pinus ldngifolia, camphora officinarum, Apium graveolens, Cedrus deodara, Eucalyptus globulus and Melaleuca leucadendron. This composition now designated as composition 2, which has been evaluated in the treatment of acute inflammation caused by carrageenin in rats.
Single injection of 0.1 ml of Carrageenin produced a significant increase in paw volume as compared to control after 2nd , 3rd and 4th hours.
The protective effect, as calculated by percentage decrease in the paw volume, was found to be significant with composition 2 as compared to carrageenin control. Composition 2 has been found to decrease in. paw volume by 12%, 45% and 46% at 2nd, 3rd, and 4th hour respectively after single application. The results demonstrating the efficacy of herbal composition on the protection of inflammation has been shown inTable:"4.
Table: 4

Model: Carrageenin (0.1 ml of 1%w/v) induced inflammation in right paw
% Decrease in Paw volume (right paw) as compared to control
Group 2 hour 3 hour 4 hour
Composition 1 51.534 57.468 42.227
Composition 2 12.368 44.905 46.219
Example 3:
Evaluation of the composition derived from the process of the present invention comprises of extract of Cissus Quadrangularis or Costus Speciosus in base oil. This composition now designated as composition 3 and composition 4, which has been evaluated in the treatment of acute inflammation caused by carrageenin in rats.
The composition containing Cissus Quadrangularis (Composition 3) produced a decrease in swelling by 14% at 2nd hour, 30% at 3rd hour and effect decreased to 13 % at 4th hour. The composition containing Costus Speciosus (Composition 4) produced a decrease in swelling by 18% at 2nd hour and 23-26% at 3rd and 4th hour. Thus, the composition containing only Cissus Quadrangularis or Costus Speciosus

(Composition 3 and 4) produced a maximum of 30% decrease with composition 3 and a maximum of 26% decrease with composition 4 in ,paw volume as compared to carrageenin-induced acute inflammation. This effect' is less than the composition 1 which contains both Cissus Quadrangularis and Costus Speciosus. So, the results indicate that the costus and cissus,:when combined together gives synergistic effect. The results demonstrating the efficacy of herbal compositions 3 and 4 on the protection of carrageenin-induced inflammation in rats have been shown in Table: 5.
Table: 5

Model: Carrageenin (0.1 ml of 1%w/v) induced inflammation in right paw
% Decrease in Paw volume (right paw) as compared to control
Group 2 hour 3 hour 4 hour
Composition 3 14.683 30.117 13.687
Composition 4 18.977, 23.679 26.735
Example 4
Removal of Cissus Quadrangularis and Costus Speciosus both or Cissus Quadrangularis or Costus Speciosus from the process for preparing composition 2 has also been evaluated. These compositions are now designated as composition 5, composition 6 and composition 7 respectively, which have been evaluated in the treatment of acute inflammation caused by carrageenin in rats.
The removal of cissus or costus, as in composition 6 and 7, from composition 2 described above leads to a drastic reduction in the activity, however, removals of both the ingredients provide a significant decrease in a paw volume on 2" and 3r hour as compared to carrageenin control as in composition 5. Composition 6 and 7 infact, produced a maximum activity of 15% and 19% respectively. However, composition 5 decreased the paw volume by 44% on 2nd hour and 37% at 3rd hour that is significantly different as compared to carrageenin control. These results also confirm the essentiality of Costus and Cissus in the composition and also the synergistic action of both the ingredients. The results demonstrating the effect of herbal compositions 5, 6 and 7 on the protection of arthritis have been shown in Table: 6.
Table: 6

Model: Carrageenin (0.1 ml of 1%w/v) induced inflammation in right paw
% Decrease in Paw volume (right paw) as compared to control
Group 2 hour 3 hour 4 hour
Composition 5 44.014 37.261 23.164
Composition 6 9.893 15.343 10.241
Composition 7 5.568 17.861 19.055

HUMAN CLINICAL TRIALS:
. A placebo controlled clinical trial was conducted on 88..patients with osteoarthritis with the composition derived from the present invention.
Patients were randomly divided in to two groups. The first group (the treatment group) consisted of 45 patients while there were 43 patients in second group (control group) In the treatment group, there were 22 males and 23 females. In control group, there were 18 males and 25 females. The patients within these two groups had no significant differences in the severity of illness.
Treatment:
Patients in the treatment group applied locally twice a day of the composition derived from the process of the present invention and patients in the control group applied placebo oil twice a day. The length of each treatment was 4 weeks. The clinical parameters were obtained before inclusion in the study and after 4 weeks of treatment.
Observations: Following observations were made to find out efficacy:

♦ Pain at rest
♦ Pain on the "use of joints
♦ Joint swelling
♦ Joint tenderness
♦ Range of movements
♦ 50 ft. walking time
♦ Duration of joint stiffness
♦ ARA functional class
♦ Visual analogue
Results:
All the parameters listed above were compared with the placebo for all the patients before and after 4 week of receiving the treatment. Pain at rest was reduced to score 1.3 from 2.5 with greater relief with the treatment as compared to placebo. Pain on the use of joints also found greater relief with the treatment as compared to placebo. Joint swelling and joint tenderness reduced better with the treatment as compared to placebo., 50 ft. walking time was also reduced by 10 minutes with the treatment as compared to 5 minutes with the placebo. The score for ARA functional class has been reduced to 1.2 from 2.4 with the treatment as compared to 1.8 from 2.5 in placebo group. Visual analogue score also improved significantly following treatment as compared to placebo. Thus the treatment of herbal composition gives better relief as compared to placebo group of patients.

Toxicity and side effect observation:
All 88 patients showed no local adverse effect during the treatment period and showed no skin reaction after the treatments.
Summary:
The study demonstrated that treatment group had significantly improved all clinical
parameters except range of movement and duration of joint stiffness as compared to
placebo. According to this study, the composition derived from the process of present
invention provide efficacy for the treatment of rheumatoid arthritis, spondylitis,
osteoarthritis, and sciatica.