PROFORMA FOR REGISTRATION OF SUBJECT FOR

DISSERTATION

  1. Name and address of the : Mr.T.NAGARJAN

Candidate S/O Naguru,

Gaddistreet,Rajampeta.

Kadapa (Dt).

AndhraPradesh.

2. Name of the guide : Mr. SYED MANSOOR AHAMED

M. Pharm, (PhD).

Assistant Professor,

Department of Pharmacology

  1. Name of the institution : SreeSiddagangaCollege of Pharmacy,

B.H.Road,Tumkur- 572 102.

  1. Course of the study : Master of Pharmacy
  1. Department : Pharmacology
  1. Date of admission: June - 2008
  1. Name of the topic : EVALUATION OF ANTIABORTIFACIENT

ACTIVITY OF NANONUT-9 FORMULATION

BRIEF RESUME OF THE INTENDED WORK

1. INTRODUCTION1:

Spontaneous abortion is noninduced embryonic or fetal death or passage of products of conception before the 20th week of pregnancy. Threatened abortion is vaginal bleeding occurring during this time frame and indicating that spontaneous abortion may occur. Diagnosis is by clinical criteria and ultrasonography. Treatment is usually with bed rest for threatened abortion and, if spontaneous abortion has occurred or appears unavoidable, uterine evacuation.Death of the fetus or passage of products of conception (fetus and placenta) before 20 week of pregnancy is considered abortion. Fetal death after 20 week is considered late fetal death and, with delivery, is considered a stillbirth. Passage of a live fetus between 20 and 37 week is considered preterm delivery.Abortions may be classified as early or late, spontaneous or induced for therapeutic or elective reasons.

Classification of abortion:

Type Definition

Early :Abortion before 12 weekgestation

Late :Abortion between 12 and 20 week gestation

Spontaneous :Abortion that is not induced

Induced :Termination of pregnancy for medical or elective reasons

Therapeutic :Termination of pregnancy to preserve the woman's life or health, or

because of fetal death or malformations incompatible with life

Threatened :Vaginal bleeding occurring before 20 week gestation without cervical

dilation, which indicates that spontaneous abortion may occur

Inevitable :Vaginal bleeding or rupture of the membranes accompanied by dilation of

the cervix

Incomplete :Expulsion of some products of conception

Complete :Expulsion of all products of conception

Recurrent : Two or more consecutive spontaneous abortions

(habitual)

Missed :Undetected death of embryo or fetus that is not expelled and causes no

bleeding (ie, blighted ovum, anembryonic pregnancy, or fetal demise)

Septic :Serious infection of the uterine contents during or shortly before or after

an abortion

About 20 to 30% of women with confirmed pregnancies bleed during the 1st 20 week of pregnancy;half of these women spontaneously abort. Thus, incidence of spontaneous abortion is about 10 to 15% in confirmed pregnancies. Incidence in all pregnancies is probably higher because some very early abortions are mistaken for a late menstrual period.

In recent decades there is a large increase in spontaneous abortion; it is mainly prevalent in young couples. In cities, estrogens such as diethyl stilbesterol and other synthetic derivatives are being used as antiabortifacient agents. These are orally active highly synthetic compounds which affect the reproductive development of the female fetus. Hence some of these agents are banned in many countries. The safety of other drugs is being used is doubtful and needs further investigation. However, a large number of plants are being used in India traditionally to prevent abortion. So far none of these plants are scientifically investigated for the folklore claim.

Nanonut-9TM is a formulation 9 highly acclaimed Ayurvedic herbs of nutritional and rasayana (antiageing, immunomodulatory, antioxidant, antiglycosylation) value2. They areAsparagus racemosus(Shatvari), Ipomoea digitata(Vidari), Trapa bispinosa(Shringatak), Embelica officinalis(Amlaki), Glycerrhiza glabra(Yashti madhu), Sida cardifolia(Bala), Withania somnifera(Ashwagandha), Hemidesmus indicus(Anantmool sariva), Tribulus terrestris(Gokhru). It cleanses blood, improves circulation, reduces blood pressure, provides high quality proteins, vitamins, and minerals for increasing fetus weight, regulates harmones and prevents threatened abortion. It is used in pregnancy and lactation for prevention of pre- eclampsia, IUGR (Intra Uterine Growth Restriction), threatened abortion, and used as a nutritional supplement. It is found to contain 15 amino acids in both free and hydrolysate form, 9 of them are essential. Studies on prevention of oxidative damage on rat liver cell mitochondria reveals that Nanonut-9is an excellent antioxidant comparible to vitamin-E2. An HPTLC study revealed the presence of gama linolenic acid in it2. The formulation is based on the experience of herbal practitioners. However so far, the antiabortifacient activity of Nanonut-9 is not proved scientifically. Hence in the present study we are interested in screening the formulation for its antiabortifacient activity in rats.

2. NEED FOR THE STUDY3:

Miscarriage or spontaneous abortion is the natural or spontaneous end of a pregnancy at a stage where the embryo or fetus is incapable of surviving, generally defined in humans at prior to 20 weeks of gestation. Miscarriage is the most common complication of early pregnancy. The medical term "spontaneous abortion" is used in reference to miscarriages because the medical term "abortion" refers to any terminated pregnancy, deliberately induced or spontaneous, although in common parlance it refers specifically to active termination of pregnancy.

Determining the prevalence of miscarriage is difficult. Many miscarriages happen very early in the pregnancy, before a woman may know she is pregnant. Treatment of women with miscarriage at home means medical statistics on miscarriage miss many cases. Prospective studies using very sensitive early pregnancy tests have found that 25% of pregnancies are miscarried by the sixth week LMP (since the woman's Last Menstrual Period). Clinical miscarriages (those occurring after the sixth week LMP) occur in 8% of pregnancies.The risk of miscarriage decreases sharply after the 10th week LMP, i.e. when the fetal stage begins. The loss rate between 8.5 weeks LMP and birth is about two percent; loss is “virtually complete by the end of the embryonic period."

Although a woman physically recovers from a miscarriage quickly, psychological recovery for parents in general can take a long time. People differ a lot in this regard, some are 'over it' after a few months, others take more than a year. Still others may feel relief or other less negative emotions.For those who do go through a process of grief, it is often as if the baby had been born but died. How short a time the fetus lived in the womb may not matter for the feeling of loss. From the moment pregnancy is discovered, the parents can start to bond with the unborn child. When the child turns out not to be viable, dreams, fantasies and plans for the future are disturbed roughly.Besides the feeling of loss, a lack of understanding by others is often important. People who have not experienced a miscarriage themselves may find it hard to empathize with what has occurred and how upsetting it may be. This may lead to unrealistic expectations of the parent’s recovery. The pregnancy and miscarriage are hardly mentioned anymore in conversation, often because the subject is too painful. This can make the woman feel particularly isolated.Interaction with pregnant women and a newborn child is often also painful for parents who have experienced miscarriage. Sometimes this makes interaction with friends, acquaintances and family very difficult.

Since there are no safe antiabortifacient drugs available, there is a need to find out antiabortifacient agents from plants. A large number of such plants are available in Ayurveda, which if validated scientifically helps a lot to the society. So far no antiabortifacient activity has been carried out on any plants. Hence the research work is of prime importance.

3. REVIEW OF LITERATURE:

1. Asperagous racemosus4:

Family: Liliaceae

Part used: Tuberous roots

Chemical constituents:The main constituents are sarsasapogenin, glucose, and rhamnose.

Ethnomedical Uses: Nutritive, galactogogue, aphrodisiac and antioxytocic activities. Reported uses: It is reported as hypolipidemic, antioxidant activities5andpossess anabolic properties viz. growth promotion.

2. Ipomoea digitata4:

Family: Convulvulaceae

Part used: Dried root

Chemical constituents:The main constituents are resins, sugars, and principally starch. Ethnomedical Uses:Aphrodisiac, lactogogue, mucilaginous and moderating menstrual discharge. Reported uses:It is reported as galactogogues, antihepatotoxic and immunomodulatory activities6.

.

3. Trapa bispinosa4:

Family: Onagraceae

Part used: Fruit

Chemical constituents: The main constituents are manganese and starch. Ethnomedical Uses:Nervine tonic, nutritive, general debility and seminal weakness.

Reported uses:It is reported as used in cytotoxic effects7 and treating indigestion especially in aesthetic children.

4.Embelica officinalis4:

Family: Euphorbiaceae

Part used: Fruit

Chemical constituents:The main constituents are vitamin-C, tannins, minerals and pectin. Ethnomedical Uses:Rejuvenating and nutritive tonic, improves all kinds of debility. Reported uses8-10:It is reported as antioxidant activity,gastro protectiveand incardiac effects.

5.Glycerrhiza glabra4:

Family: Papilionaceae

Part used: Roots

Chemical constituents: The main constituents are glycyrrhizins, asparagines, sugar, acid resin, phosphoric, sulphuric and maleic acids, cadium and magnesium salts. Ethnomedical Uses:Astringent, nervine tonic, relieving pain and discomfortand uterine complaints. Reported uses11:It is reported as used in polycystic ovary syndrome andestrogeniceffects.

6. Sida cardifolia4: Family: Malvaceaea Part used: Roots Chemical constituents:The main constituents are alkaloids (ephedrine), phytosterols, resin acids and fatty oils. Ethnomedical Uses:Nervine tonic, haematuriaand cardiac tonic. Reported uses12: It is reported as analgesic activity, anti-inflammatory and hypoglycaemic activities.

7. Withania somnifera4:

Family:Solanaceae

Part used: Roots

Chemical constituents:The main constituents are alkaloids (withanine), steroidal lactones(withanolides) and 16beta-acetoxy-17(20)-ene. Ethnomedical Uses:Health restorative to pregnant, rejuvenating drug and as aphrodisiac. Reported uses:It is reported as higher sexual performance, antistress, antioxidant, immune modulating13 and rejuvenating properties14.

8. Hemidesmusindicus4:

Family:Asclepiadaceae

Part used: Roots

Chemical constituents:The main constituents are coumarin, hemidesmine, smilasperic acid,enzymes, essential oils and a saponin. Ethnomedical Uses:Nutritional disorders, sexual debility and all kinds of disease caused by vitiated blood. Reported uses15:It is reported as antioxidant activity,antithrombotic activity,antiplatelet aggregation and anticoagulant activity.

9. Tribulus terrestris4:

Family:Zygophyllaceae

Part used: Dried spiny fruit

Chemical constituents:The main constituents are alkaloids, resins, fat and mineral matter (Nitrates). Ethnomedical Uses:To ensure fecundity, impotence, seminal debility and as aphrodisiac. Reported uses:It is reported as antihypertensive16 and asaphrodisiac activity17.

4. OBJECTIVE OF STUDY:n4

To evaluate the antiabortifacient activity of Nanonut-9TMformulation.

5. MATERIALS AND METHODS:

5.1. Collection of formulation: TheNanonut-9 capsuleswill be collected from Phytocastle industry which is present in Bangalore.

5.2. Preparation of suspension:A suspension of formulation will be prepared in distilled water suspended with Tween-80.

5.3. Animals: Animals used in the experiment will be colony bred wristar strain female albino rats weighing 150-200g. All the animals will be maintained under controlled standard husbandry conditions with food and water ad libitum.

5.4. Selection of dose: 100 mg/kg b.w and 200 mg/kg b.w.

5.5. Route of administration: Oral route

5.6. Method: Proven fertile female rats will be selected and 10 will be used for each control and experimental group. Rats having regular estrous cycle alone taken for the studies and mated with proven fertile males in the ratio of 2:1 in the estrous phase of the estrous cycle. The presence of spermatozoa in vaginal smear will be taken as day 1 of pregnancy.On day 7 of pregnancy ultrasonography scanning will be done and the number of implantation sites will be counted. The following treatments will be made on day 8 to day 14 of pregnancy to different groups of rats containing 10 in each group. The animals will be observed for any vaginal bleeding after administration of the extracts and standard.

On day 16 of the pregnancy ultrasonography scanning will be carried out and the number of implantation sites in each animal will be noted. If implantation sites were present, the animals will be allowed to grow for full term. The number of litters born were noted at the end of gestation and checked for signs of gross malformation. The increase in the number of implantation sites in the treated group indicates antiabortifacient activity.

6. EXPERIMENTAL DESIGN:

S.no / Drug / Dose
(Oral) / No.of animals / Parameters
1. / Control / ………. / 10 / 1. Number of
implantation sites
2. Number of litters
2. / Standard drug ( Mifepristone ) / 20 mg/kg / 10
3. / Nanonut-9 / 100 mg/kg / 10
4. / Nanonut-9 / 200 mg/kg / 10
5. / Mifepristone+ Nanonut-9 / (20+100)mg/kg / 10
6. / Mifepristone + Nanonut-9 / (20+200)mg/kg / 10

7. BIBLIOGRAPHY:

  1. Nanonut-9TM formulation brochure Phytocastle,Byadarhalli, Magadi main road, Bangalore-560091.
  2. Nadkarni KM, Nadkarni AK.Indian materia medica.Bombay; Popular Prakashan Pvt.Ltd.1994;Vol1.
  3. Visavadiya NP, Narasimhacharya RL. Hypolipidemic and antioxidant activities of

Asparagus racemosus in hypercholesteremic rats.Ind J Pharmacol 2005; 37:376-80.

Year=2003; volume=57; issue=9; spage=408; epage=14; aulast=Goyal.

  1. Kosuge T, Yokota M, Sugiyama K, Okamoto A,Saito M, Yamamoto T.Studies on Chinese medicines used for cancer. III. Cytotoxic constituent against HeLa cells in the fruit of Trapa bispinosa Roxb 1986; 106: 183-85.
  2. Scartezzini P, Antognoni F, Raggi MA, Poli F, Sabbioni C.Vitamin C content and antioxidant activity of the fruit and of the Ayurvedic preparation of Emblica officinalis. J Ethnopharmacol2006; 104: 113-18.
  3. Al-Rehaily AJ, Al-Howiriny TA, Al-Sohaibani MO, Rafatullah S.Gastroprotectiveeffects of 'Amla' Emblica officinalis on in vivo test models in rats. Phytomedicine 2002; 9: 515-22.
  4. Rajak S, BanerjeeSK, Sood S. Emblica officinalis causes myocardial adaptation and

protects against oxidative stress in ischemic-reperfusion injury in rats. Phytother Res

2004; 18:54-60.

  1. Armanini D, Castello R, Scaroni C. Treatment of polycystic ovary syndrome with

spironolactone plus Licorice. Eur J Obstet Gynecol Reprod Biol. 2007; 131: 61-67.

  1. Kanth VR, Diwan PV.Analgesic, anti-inflammatory and hypoglycemic activities of

Sida cardifolia.Phytother Res 1999;13:75-77.

  1. Ziauddin M. Studies on the Immunomodulatory Effects of Ashwagandha. J Ethnopharmacol 1996; 50:69-76.
  1. Jain S, ShuklaSD, Sharma K, Bhatnagar M. Neuroprotective Effects of Withaniasomnifera Dunn. in hippocampal sub-regions of female albino rat. Phytother Res2001; 15:544-48. .
  2. Mary NK, Achuthan CR, BabuBH, Padikkala.. In vitro antioxidant and

antithrombotic activity of Hemidesmus indicusJEthanopharmacol2003; 87:187-91.

  1. Sharifi AM, Darabi R, Akbarloo N.Study of antihypertensive mechanism of Tribulusterrestris in 2K1C hypertensive rats: role of tissue ACE activity. Life Sci. 2003; 73: 2963-71.
  2. Gauthaman K, Adaikan PG, Prasad RN. Aphrodisiac properties of Tribulus terrestris

extract (Protodioscin) in normal and castrated rats. Life Sci2002; 71:1385-96.

8. CLEARENCE FROM INSTITUTIONAL ETHICAL COMMITTEE:

The study is cleared from Animal Ethical Committee of the Institution.

(Approval no: 62/2008-09, Dated: 17-11-2008) Enclosed Copy. Annexure-1

SIGNATURE OF THE CANDIDATE-

(T.NAGARJAN)

9. REMARK OF THE GUIDE:

The present study will be carried out under my supervision and Guidance.

SIGNATURE-

(SYED MANSOOR AHAMED)

10.REMARK OF HEAD OF THE DEPARTMENT:

The present work can be carried out in our department; the protocol is approved by me.

SIGNATURE-

(Dr. SHIVA KUMAR SWAMY)

11. REMARK OF THE CHAIRMAN AND PRINCIPAL:

The above-mentioned information is correct and recommended for approval.

SIGNATURE-

(Dr. S. BADAMI)

1