“COMPARATIVE ASSESSMENT OF ANTI-OSTEOARTHRITIC ACTIVITY OF TRAYODASHANGA GUGGULU(A POLY HERBAL FORMULATION) AND
GLUCOSAMINE SULPHATE”
SYNOPSIS FOR
M. PHARM DISSERTATION
SUBMITTED TO
RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BENGALURU
SUBMITTED BY
NETHRAVATHI
I M. PHARM
DEPARTMENT OF PHARMACOLOGY
GOVERNMENTCOLLEGE OF PHARMACY
BENGALURU – 560 027
(2013-2014)
UNDER THE GUIDANCE OF
Dr. S. RAMACHANDRA SETTY
M.pharm PhD
PROFESSOR& HEAD
DEPARTMENT OF PHARMACOLOGY
GOVERNMENT COLLEGE OF PHARMACY
BENGALURU - 560 027
RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BENGALURU
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECT FOR
M PHARM PART-I DISSERTATION
1.0 / NAME AND ADDRESS OF THE CANDIDATE. / NETHRAVATHIPRESENT ADDRESS
C/O Mahadeva, #64/4,
Channappa layout,
Roopenaagrahara,
Bangalore 560 068.
PERMANENT ADDRESS
D/O Adinarayana Reddy,
Devgana halli, bychapura post,
Gauribidanur.(Tq). Chikkaballapur dist.
2.0 / NAME OF THE GUIDE / DR. S RAMACHANDRA SETTY
M.pharm, Ph.D.
Professor and HOD,
Dept. of pharmacology.
3.0 / NAME OF THE INSTITUTION. / Government College of
Pharmacy Bangalore 560 027.
4.0 / COURSE OF STUDY AND SUBJECT. / M.pharm
Pharmacology.
5.0 / DATE OF ADMISSION TO COURSE. / 01thAugust, 2013
6.0 / “COMPARATIVE ASSESSMENT OF ANTI-OSTEOARTHRITIC ACTIVITY OF TRAYODASHANGA GUGGULU(A POLY HERBAL FORMULATION) AND GLUCOSAMINE SULPHATE”
6.1. NEED FOR THE STUDY:
Living with pain is the worst tragedy that can happen to any living being. Vatadosha is considered to play a major role in maintaining samyavastha of tridosha1. Vatadosha is also responsible for the vitiation of other doshadhatu and mala. SandhiGataVata (SGV) is explained in ayurveda under vatavyadhi the concept of Gata vata2is explained where in group of symptoms seen in vatadosha when it afflicts dhatu, upadhatu and Sandhi. SandhiGataVata is characterized by pain, swelling and restricted movement of joints3. These explanations f Ayurveda and the symptoms described show a lot of similarity with osteoarthritis of contemporary science.
Osteoarthritis(OA): It is the most common form of arthritis. Cartilage present (connective tissue) between the bones gradually wastes away (degenerates), and lead to painful rubbing of bone on bone in the joints. It may cause joints to fall out of their natural positions (misalignment), deformity in the bone structure, and most frequently affected joints are in the hands, spine, knees and hips.4
Arthritis shares the symptoms of pain, stiffness, and swelling of one or more joints. OA involves a wear-and-tear degeneration of cartilage in the joint space with only minimal inflammation.5
Arthritis is predominantly a disease of theelderly, but children/adults can also be affected by the disease. Approximately 350 million people worldwide suffer from arthritis.Nearly 40 million persons in the United State are affected by arthritis, including over a quarter million childrenand physical disability in the older people. Today 35 million people (13% of the US population) are 65 and older, and more than half of them have radiological evidence of OA in at least one joint4. Prevalence of OA after age 65 is about 60% in men and 70% in women.6
At present, therapies available to treat OA are limited. Most current treatments are designed only to relieve pain and reduce/prevent the disability caused by bone and cartilage degeneration. Drug therapies target the symptoms but not the cause of this disease, no treatment inhibits the degenerative structural changes that are responsible for its progression and these treatments are less satisfactory and may lead to side effects such as in case of following drugs4.
In addition several drugs adopted in the treatment of OA also have severe side effects. For example;
NSAIDS: damage to upper and lower gastrointestinal tract
Corticosteroids: damage cartilage.
Therefore there is a need to develop new strategies to treat these disorders. Hence, the goal of present study is to integrate allopathic system of medicine with Ayurvedic (polyherbal formulation) so as to demonstrate the utility of such treatment strategy in preventing, retarding the progression of cartilage degeneration and enhancing the safety of the treatment. OA is caused due to vitiation of vata (Air) as mentioned in Ayurveda. Joints have a lubricating substance which facilitates the appropriate mobility of these joints. The aging process and other degeneration gradually dries the lubricating substance. This condition is termed as "SandighataVata' (osteo arthritis). This needs the replenishment of the lubrication of the affected joints. Here also TraydoshangGuggulu claimed to eradicate the Vata (Air) dosha (Vitiation).7
Glucosamine is an aminosaccharide, precursor forglycosaminoglycans, and glycosaminoglycans are a major component ofjoint cartilage, supplemental glucosamine may influence cartilage structure, and so may apply to alleviation of arthritis4. Its use as a therapy for osteoarthritis appears safe. So, combination of these two may synergize the effect of each other and enhance the efficacy of the treatment.It’s marketed as glucosamine sulphate. Glucosamine is marketed to support the structure and function of joints and the marketing is targeted to people suffering fromosteoarthritis10.In the present study 3 experimental models of osteoarthritis will be adapted to assess the study hypothesis. They are
1) Formaldehyde induced OA in rats(which acts by chronic inflammation )15, 16
2) Monosodium iodoacetate induced osteoarthritis in rats(which act by disturbing chondrocyte metabolism)14
3) Papain induced osteoarthritis in rats(which act by matrix degranulation of articular cartilage using enzymatic means)17, 18
TrayodashangaGuggulu is mainly known to bring about reduction of vata (Air) Dosha (vitiation) within the body.
As per Ayurvedic texts and references the vata (Air) Dosha (vitiation) produces problems in the body . Once it is released out of its main position in the body this vata (Air) will move throughout the body producing further problems by carrying the other two doshas (vitiation) i.e. Pita (fire/Agni) and Kapha (water) along with it from their positions and spreading out into the body producing problems. So this herbal preparation brings about the cure of the problems by confining vata (Air) in its original position.8Glucosamine is a precursor forglycosaminoglycans, and glycosaminoglycans are a major component ofjoint cartilage, supplemental glucosamine may influence cartilage structure, and so may apply to alleviation of arthritis.11
Hence the study of the drug glucosamine sulphate and polyherbal formulation TrayodashangaGuggulu act by two different mechanisms, which can prevent cartilage degeneration, the combination of them may be highly useful.
Therefore the study is needed and justifiable.
6.2 .REVIEW OF THE LITERATURE:
CONTENTS OF TRAYODASHANGA GUGGULU7,8:
NAME OF THE PLANT / QUANTITY1. Abha (Babbula) (acacia nilotica) (Stem. Bark) / 1 Part
2. Ashwagandha (Withaniasomnifera) (Root) / 1 Part
3. Hapusha (JuniperusCommunis) (Fruit) / 1 Part
4. Guduchi (TinosporaCordifolia) (Stem) / 1 Part
5. shatavari (asparagus racemosus) (tuberous Root) / 1 Part
6. Gokshura (Tribulusterrestris) (Fruit) / 1 Part
7. Vrddhadaru (argyreiaspeciosa) (Root) / 1 Part
8. Rasna (alpiniaofficinarum) (Root/Leaf) / 1 Part
9. shatahva (Anthumsowa) (Fruit) / 1 Part
10. Sati (hedychiumspicatum) (rhizome) / 1 Part
11.Yavani (carumcopticum) (Fruit) / 1 Part
12. Sunthi (Zinger officinalis) (rhizome) / 1 Part
13. SuddhaGuggulu (commiphoramukul) (Extracted) / 12 parts
14.Goghuta (ghee) / 1 Part
TrayodashangaGuggul is a very famous Ayurvedic medicine. It is in tablet form. It is also known asTrayodashangaGuggul tablets, TrayodashangaGuggulu etc. This tablet has Guggulu (commiphoramukul) as its base. It is widely used in the Ayurvedic treatment of joint pain related disorders. Here, Guggulu is treated with 13 ingredients so the name Trayodashanga(Trayodashameans thirteen).7The herbalformulation is used to promote appetite and digestion, increase the number of red blood cells, and aid in removal of undesirable fat in the body. Further Ayurvedic texts describes that this herbal preparation is very beneficial in many types of VataDosha diseases. This is also claimed to be beneficial in Vatashoola. There is a strange claim that regular use of by a limping or crippled person, bring about reduction in their limp as well as cripple. This can even care internal organs of the body as a mother cares for her child. It promotes the internal cleansing, reduce conditions of stagnation and excess, and improve digestion and assimilation of nutrients.Trivrut(Operculinaturpethum).7
Guduchi is one of the most versatile rejuvenative herbs, it boosts the immunity, promote longitivity. In this formulation it enhances the action of Guggulu and works well to impart additive effects. It is bitter, pungent and astringent in taste, sweet in post digestive effect and hot potency. It alleviates all the tridosas. Except pungent post digestive effect, Guggulu possess the same properties as that of guduchi. Guggulu is one of the best rewarding drug for the vata diseases. The combination of both these herbs becomes a potent remedy for various vata disorders.
Ashwagandha;Plant pacifies vitiatedvata (AIR), nerve weakness, impotency, arthritis, the roots are aphrodisiac and stimulant widely used in rejuvenativeAyurvedic formulation. Plant has immuno stimulant action.
Shatavari - Asparagus Racemosus; Plant pacifies vitiated pitta (Agni/fire), vata (air), leucorrhoea, inflammation, nervous debility.
Gokhru - TribulusTerrestris; Plant pacifies vitiated vata, Pitta, urinary retention, UTI (urinary tract infection), Sexual weakness, anemia, general weakness, constipation
Rasna - PlucheaLanceolata: The Herb is very much useful in rheumatism, swelling of joints in arthritis, neurological disorders. It is also used as antipyretic, analgesic, laxative and nervine toxic.. Regular use of Rasna delays the signs of ageing.
TrayodashangaGuggul benefits: It is widely used in the Ayurvedic treatment for low back pain, sciatica, arm pain, locked jaw, knee joint pain, foot pain, arthritis pain and myalgia.19
1. Trayodashanga is one of the best herbal remedy for various types of VataDosha diseases. This is very well known for its ability to bring about the cure ofmost chronic VataDosha diseases.
2. Trayodashanga is also beneficial in Vatashoola.
3. Trayodashanga is also known to bring about the treatment of paralysis. This is brought about by its ability to cure the excessive Vata in the body and since paralysis is resulted due to the excessive Vata in the body so it is easily cured by this herbal preparation. So it is best for joint pain herbal treatment
4. Trayodashanga is also beneficial in treatment of arthritis.
5. Trayodashanga is also used for the treatment of hemiplegia, paraplegia etc.
6. Trayodashanga is known for its ability to bring about relief in Sciatica pain.7,8
Important Therapeutic Uses of Trayodashanga Guggulu8;
· KatiGraha(Stiffness inlumbo-sacral region),
· Grudhrasi(Sciatica),
· Hanugraha(Lock jaw),
· Bahushula(Pain in arm),
· JanuStabdhata(Stiffness of the knee),
· Asthivata(Bone diseases due toVatadosha),
· Majjavata(Bone marrow disorder),
· Snayuvata(Inflammation of ligaments),
· HruthGraha(Cardiac failure),
· Vatakapharoga(Disease due toVataKaphadosha),
· Yonidosha(Disorder of female genital tract),
· AsthiBhagna(Bone fracture),
· Vidradhi(Abscess),
· KhanjaVata(Limping)
Glucosamine sulphate: (3R, 4R, 5S)-3-Amino-6-(hydroxymethyl) oxane-2, 4, 5-triol.
It is an aminosaccharide, acting as a preferred substrate for the biosynthesis of glycosaminoglycans chains and subsequently for the production of aggrecan and other Proteoglycans of cartilage4.
Glucosamine sulphate&Chondroitinsulphate marked as nutritional supplements in US12 and considered to have beneficial effects on cartilage. It tends to replace degenerated cartilage9.
Clinical studies have consistently reported that glucosamine appears safe. However, a recentUniversity Lavalstudy shows that people taking glucosamine tend to go beyond recommended guidelines, as they do not feel any positive effects from the dietary supplement. Beyond recommended dosages, researchers found in preliminary studies that glucosamine may damage pancreatic cells, possibly increasing the risk of developing diabetes. Adverse effects, which are usually mild and infrequent, include stomach upset, constipation, diarrhea, headache and rash4.
6.3 OBJECTIVE OF THE STUDY:
Ø To evaluate the anti-osteoarthritic activity of polyherbal formulation TrayodashangaGuggulu and glucosamine sulphate against monosodium iodoacetate, formalin, and papaininduced OA activity in rats.
Ø To determine the beneficial combination of TrayodashangaGuggulu and glucosamine sulphate treatment in monosodium iodo acetate, formalin and papain induced OA in rats.
Ø To study whether the polyherbal formulation can overcome the side effects caused by standard drugs.
MATERIALS AND METHODS
6.4 SOURCE OF DATA: data will be generated by performing experiments on animals. The standard information from collect from various journals, standard text books in library of Govt. College of pharmacy, Indian institute of science, RGUHS digital library and from various standard websites.
Web sites: www.pubmed.com
www.google.com
www.wikipedia.org
ANIMALS
Adult rats weighing between 200-250g of either sex will be used in the study. The animals will be kept under standard conditions at ambient temp of 25±20 C with 12hr light/dark cycle, with food and water. Animals will be acclimatized to laboratory condition before the test.
TOTAL ANIMALS REQUIRED
Total animals used in acute toxicity studies=20mice
Animals used in each group= 06 rats
Total rats use =90 rats
ACUTE TOXICITY STUDIES:
Determination of LD50 of TrayodashangaGuggulu as per OECD guidelines, the guideline described by OECD will be adapted for the determination of LD50 on adult female mice. Female mice will be used as they are more sensitive to toxicity compared to male mice, hence female mice are taken for acute toxicity study and1/10th of LD50 dose will be taken for further studies.
EXPERIMENTAL DESIGN
1) FORMALDEHYDE INDUCED OSTEOARTHRITIC MODEL15,16:
Total 30 rats of either sex will be randomly divided into 5 groups of 6 rats each and treated for
3days as follows;
Group 1: receives only vehicle (p.o.) for 3 days.
Group 2: receives formaldehyde solution 0.1 mL of 2 %( i.a)+normal saline (p.o) for 3 days.
Group 3: receives formaldehyde solution 0.1 mL of 2%(i.a)+ TrayodashangaGuggulu
Group 4: receives formaldehyde solution 0.1 mL of 2%(i.a)+glucosamine sulphate (p.o.)
Group 5: receives normal saline(i.p.)+ TrayodashangaGuggulu and glucosamine sulphate(p.o.)
On day one, arthritis was induced by injecting 0.1 mL of 2.0% formaldehyde solution into the right hind, and this procedure was repeated on day 3, rats of all groups, except the negative control, were administered with the respective treatments (vehicle, TrayodashangaGuggulu, glucosamine sulphate). One hour later, the diameter of the paw and that of the right ankle were measured on day one, before the induction of the damage, and after 10 days on treatment. Measurements were done by determining the perimeter, using a tied thread around the surface and a graded rule. The anti-arthritic activity assesses as the anti- inflammatory response, was calculated as follows: %= (1-T/C) x 100, where T and C are the mean diameters of the treated and positive control groups, respectively.