1

1. Introduction

Diabetes is a metabolic disorder characterized by hyperglycemia, glycosuria,

hyperlipaemia, negative nitrogen balance and sometimes ketonaemia. A widespread

pathological change is thickening of capillary basement membrane, increase in vessel

wall matrix and cellular proliferation resulting in vascular complications like lumen

narrowing, early atherosclerosis, sclerosis of glomerular capillaries, retinopathy,

neuropathy and peripheral vascular insufficiency. Two major types of diabetes mellitus

are Type1: Insulin-dependent diabetes mellitus and Type2: Noninsulin-dependent

diabetes mellitus. (1)

Type-1: Insulin Dependent Diabetes Mellitus (IDDM) - In this condition, there is

no production of insulin by the pancreas and the patient is totally dependent upon

externally administered insulin. Type 1 diabetes is primarily due to autoimmune -

mediated destruction of pancreatic β-cell islets, resulting in absolute insulin

deficiency. People with type 1 diabetes must take exogenous insulin for the

survival to prevent the development of ketoacidosis. Its frequency is low related to

type 2 diabetes, which accounts for 90% of the case globally.

Type-2: Non-Insulin Dependent Diabetes Mellitus (NIDDM) - This condition is

caused by the following factors:-

a) Insufficient production of insulin from pancreas.

b) Peripheral resistance by the cells to the action of available Insulin.

c) Both a and b

Type 2 diabetes is characterized by insulin resistance and/or abnormal insulin

secretion, either of which may predominate. People of type 2 diabetes are not

dependent on exogenous insulin, but may require it for controlled blood glucose

levels if this is not achieved with diet alone or with oral hypoglycemic agents. The

epidemic diabetes relates particularly to type 2 diabetes. (2 )

The prevalence of diabetes for all age-groups worldwide was estimated to be 2.8%in

2000 and 4.4% in 2030. The total number of people with diabetes is projected to rise

from 171 million in 2000 to 366 million in 2030. The prevalence of diabetes is higher in

men than women, but there are more women with diabetes than men. The urban

2

population in developing countries is projected 2000 and 2030. The most important

demographic change to diabetes prevalence across the world appears to be the increase in

the proportion of people >65 years of age. The number of people with diabetes is

increasing due to population growth, aging, urbanization, and increasing prevalence of

obesity and physical inactivity. (3)

Table 1.1: Showing list of countries with the highest number of estimated cases of

diabetes (3)

2000 2030

Ranking country People with

diabetes(millions)

country People with

diabetes(millions)

1 India 31.7 India 79.4

2 China 20.8 China 42.3

3 U.S. 17.7 U.S. 30.3

4 Indonesia 8.4 Indonesia 21.3

5 Japan 6.8 Japan 8.9

6 Pakistan 5.2 Pakistan 13.9

7 Bangladesh 3.2 Bangladesh 11.1

8 Brazil 4.6 Brazil 11.3

The oral hypoglycemic agents/insulin are the mainstay of treatment of diabetes and are

effective in controlling hyperglycemia, but have prominent side effects & fail to

significantly alter the course of diabetic complications though development of modern

medicine resulted in the advent of modern pharmacotherapeutics including, Biguanides,

sulfonylureas and thiazolidinediones (Table 1.2), there is still a need to look for new

drugs as no drug (except strict glycemic control with insulin has been shown to modify

the course of diabetic complications).

3

Table 1.2: Oral Hypoglycemic Drugs: Allopathic formulations used for the

treatment of diabetes (1)

Drug Administration

time

Dosing schedule Possible side

effects

Comments

Glipizide Taken with meal Usually taken in

1 or 2 doses

Hypoglycemia Interact with oral

anticoagulants

Metformin Taken during or

immediately meal to

minimize

gastrointestinal side

effects

Usually taken in

1 or 3 doses

GI disturbance Should be

stopped before

surgery and

radiological

investigations

involving

contrast media

Acarbose Swallow whole with

liquid before meal or

chew with the first

few mouthfuls of

food

Usually taken in

1 or 3 doses

GI disturbance Sucrose should

not be

administered if

the patient

experience

hypoglycemia

Repaglinide Taken with meal Usually taken 3

times in a day

Hypoglycemia -

Pioglitazone Taken with meal Usually taken in

single dose

Hypoglycemia -

Glibenclamide Taken with meal Usually taken in

1 or 2 doses

Hypoglycemia Interact with oral

anticoagulants

Glimiperide Taken with meal or

15 to 30 mins.

Before meal

Usually taken in

single dose

Hypoglycemia Interact with oral

anticoagulant

Gliclazide Taken with meal Usually taken in

1 or 2 doses

Hypoglycemia Interact with oral

anticoagulant

4

Table 1.3: Ayurvedic formulation for diabetes (4)

Drug Company Ingredients

Diabecon Himalaya Gymnema sylvestre, Pterocarpus

marsupium, Glycyrrhiza glabra,

Casearia esculenta, Syzygium cumini,

Asparagus racemosus, Boerhavia

diffusa, Sphaeranthus indicus,

Tinospora cordifolia, Swertia chirata,

Tribulus terrestris, Phyllanthus

amarus, Gmelina arborea, Gossypium

herbaceum, Berberis aristata, Aloe

vera, Triphala, Commiphora wightii,

shilajeet, Momordica charantia, Piper

nigrum, Ocimum sanctum, Abutilon

indicum, Curcuma longa.

Pancreatic tonic 180 cp Ayurvedic herbal

supplement

Pterocarpus marsupium, Gymnema

sylvestre, Momordica charantia,

Syzygium cumini, Trigonella foenum

graceum, Azadirachta indica, Ficus

racemosa, Aegle marmelos,

Cinnamomum tamala

Ayurveda alternative

herbal formula to

Diabetes:

Chakrapani Ayurveda Gurmar (Gymnema sylvestre) Karela

(Momordica charantia) Pushkarmool

(Inula racemosa) Jamun Gutli

(Syzygium cumini) Neem (Azadirachta

indica) Methika (Trigonella foenum

gracecum) Guduchi (Tinospora

cordifolia)

Bitter gourd Powder Garry and Sun natural

Remedies

Bitter gourd (Momordica charantia)

5

Bitter gourd Powder Garry and Sun natural

Remedies

Bitter gourd (Momordica charantia)

Diabetes-Daily Care Nature‟s Health Supply Alpha Lipoic Acid, Cinnamon 4%

Extract, Chromax, Vanadium,

Fenugreek 50% extract, Gymnema

sylvestre 25% extract Momordica 7%

extract, Licorice Root 20% extract

Gurmar powder Garry and Sun natural

Remedies

Gurmar (Gymnema sylvestre)

Epinsulin Swastik Formulations vijaysar (Pterocarpus marsupium)

Diabecure Nature beaute santé Juglans regia, Berberis vulgaris,

Erytherea centaurium, Millefolium,

Taraxacum

Diabeta Ayurvedic cure Ayurvedic

Herbal Health Products

Gymnema sylvestre, Vinca rosea

(Periwinkle), Curcuma longa

(Turmeric), Azadirachta indica

(Neem), Pterocarpus marsupium

(Kino Tree), Momordica charantia

(Bitter Gourd), Syzygiumcumini (Black

Plum), Acacia arabica (Black Babhul),

Tinospora cordifolia , Zingiber

officinale (Ginger)

Syndrex Plethico Laboretaries Germinated Fenugreek seed extract

According to ayurveda many medicinal plants (Allium cepa, Allium sativum, Aloe

vera , Cajanus cajan , Coccinia indica , Caesalpinia bonducella ,Ficus bengalenesis ,

Gymnema sylvestre , Momordica charantia , Ocimum sanctum, Pterocarpus marsupium,

Swertia chirayita, Syzigium cumini, Tinospora cordifolia and Trigonella foenum

6

graecum.) have hypoglycemic effects & that covers the 70% global market of health

system. It suggests that Type II diabetes is a Kapha disorder, brought on by poor

digestion and accumulation of ama (toxicity) which clogs cell membranes and inhibits the

passage of glucose through the cell wall.(5)

Table 1.4: Use of traditional and other medicinal plants for antidiabetic activity

Sr.

No

Plant Sr. No. Plant

1 Acacia arabica: (Babhul) 20 Ficus bengalensis : (Indian

banyan tree)

2 Areca catechu: (Supari) 21 Gymnema sylvestre : (Gurmar

or merasingi)

3 Aegle marmelos: (Bengal

Quince, Bel or Bilva)

22 Hibiscus rosa-sinensis :

(Gudhal)

4 Allium cepa: (Onion) 23 Ipomoea batatas :

(Sakkarkand or Mitha aalu)

5 Allium sativum : (Garlic) 24 Lantana camara : (Caturang

or Ghaneri)

6 Aloe vera and Aloe barbadensis 25 Memecylon umbellatum :

(Anjali or Alli)

7 Artemisia pallens : (Davana) 26 Momordica charantia : (Bitter

gourd)

8 Azadirachta indica : (Neem) 27 Morus alba : (Shetut or Tut)

9 Biophytum sensit ivum : (Lajjalu

or Laksmana)

28 Musa sapientum : (Kela)

10 Beta vulgeris : (Chukkander) 29 Mucuna pruriens : (Kavach)

11 Brassica juncea : (Rai) 30 Murraya koeingii :

(Currypatta)

12 Bombax ceiba: (Semul) 31 Nelumbo nucifera : (Kamal)

13 Caesalpinia bonducella 32 Ocimum sanctum: (Holy basil)

7

14 Cajanus cajan : (Tuvar) 33 Picrorrhiza kurroa : (Kutaki)

15 Capparis decidua 34 Phyllanthus niruri : (Jangli

amla)

16 Coccinia indica 35 Pterocarpus marsupium :

(Vijayasar or Bijasal)

17 Citrullus colocynthis: (Badi

Indrayan or Makkal)

36 Trigonella foenum graecum :

(Fenugreek)

18 Eucalyptus globulus: (Safeda) 37 Tinospora cordifolia:

(Guduchi)

19 Eugenia jambolana : (Indian

gooseberry, jamun)

38 Momordica cymbalaria :

(Kadavanchi and Athalaki)

In the review of literature, it was found that the Cordia Dichotoma Forst. and Carissa

Carandas plant have not the used in the herbal formulation till date, the reasons behind

these antidiabetic activities of both plant are not be fully discovered.

Cordia Dichotoma Forst. f. (Boraginaceae) is commonly known as „Bhokar‟ in

Marathi. It is a small to moderate-sized deciduous tree with a short bole and spreading

crown. (6) It is an important drug for indigenous systems of medicine and has been

attributed with many medicinal properties in Ayurveda. The plant has been claimed to

cure diseases of the kidney, liver, spleen, heart, and blood. Various parts of the plants are

used as an antipyretic, for anti-anemic effect, as a remedy for impotency, and to treat

gastric pain, asthma, mouth ulcers, bronchitis, diarrhea, rheumatism, and dental caries (7,

8). Traditionally bark of the plant is reported for the treatment of ulcerative colitis (6, 7).

Present work was undertaken to find out probable phytoconstituent responsible for the

treatment of ulcerative colitis.

Carissa Carandas commonly known as Karanda belongs to family Apocynaceae.

In recent years its botanical name was changed to Carissa congesta Wight (syn. Carissa

Carandas Auct. Formerly widely known as Carissa Carandas L.) (9). Carissa Carandas is

large dichotomously branched evergreen shrub with short stem and strong thorn in pairs.

This species is a rank-growing, straggly, woody, climbing shrub, usually growing to 10

8

or 15 ft. (3–5m) high, sometimes ascending to the tops of tall trees. The plant is native

and common throughout India (10). Traditionally, whole plant and its parts were used in

the treatment of various ailments. The roots were employed as a bitter stomachic,

vermifuge and as an ingredient in the remedy for itches. The roots were reported to

contain salicylic acid and cardiac glycosides. It also contains carissone; d-glycoside of

sitosterol; glucosides of odoroside H; carindone; a terpenoid lupeol; ursolic acid and its

methyl ester; also carinol, a phenolic lignan. Fruits contains good amount of vitamin C.

The fruits, leaves and bark are rich in tannins (11).

In Ayurveda, the unripe fruits were used as an anthelmintic, astringent, appetizer,

antipyretic, antidiabetic, aphrodisiac, in biliary disorders, stomach disorders, rheumatism

and diseases of the brains (11, 12). It is useful in treatment of diarrhea, anorexia and

intermittent fevers. Fruits have also been studied for its analgesic, anti-inflammatory and

lipase 1 activities (13). It is used by tribal healers of Western Ghat region of Karnataka as

hepatoprotective and antihyperglycemic. However, no scientific data is available to

validate the folklore claim (10, 14). Keeping the above information in view, the present

study has been designed to evaluate its antidiabetic activity.

The Ayurveda system (15, 16) has defined the antidiabetic effects of the Carissa Carandas

and Cordia Dichotoma Forst.

[Meaning: Unripe and ripe both verities of Carissa Carandas are heavy in digestion and

used as the anti-thirst, Produces heat, used in anorexia and initiates the RAKT, PITTA

(heat) & KAPHA (water), Ripe fruit are sweet, used in anorexia, light in digestion and

used in the PITTA & VATA Dash disorders].

According to Ayurveda Vata known as the vatik prameha, it is further divide into four

types: vasameha, mejajamehe, lasicameha, oozumeha (15) also has known as the

medhumeha i.e. Diabetic mellitus.

9

[Meaning: karamard is described in Ayurveda as “GRAHI” substance. Grahi means

which has appetizer & digestive properties and is liquid absorber, it is known as

“Deravshosak”].

Since the two major limiting factors for molecules to pass the biological

membrane for their absorption in the blood streams mainly includes lipid solubility and

molecular sizes. There are many plant extract having excellent bioactivity in vitro but low

or less in vivo because of their poor lipid solubility and improper size of the molecule or

both which result in poor absorption and bioavailability of constituents from plant extract

are destroyed in the gastric fluids when taken orally.(18, 19, 20)

Thus bioavailability is most important factor for therapeutics & there are

bioavailability enhancement technique which aids in modifying the solubility of plant

extracts and thus improving the absorption and bioavailability. There are many types of

solubility enhancement techniques but the most common is physical modification i.e.

solid dispersions, microemulsions, eutectic mixtures and complexation etc. complexation

is the association between the two or more molecule to form a bonded/non-bonded entity

with a well-defined stoichiometry. Various complexing agents such as EDTA,

cyclodextrins and polymer have been used for the complexation. Curcumin is poorly

soluble drug hence its oral use is limited owing to its solubility. The solubility of the

curcumin was increased by the formation of the curcumin soya lecithin complex and

evaluated for the hepatoprotective activity. The in vitro permeation study of complex

showed increase in the permeability of the drug leading to greater hepatoprotective

activity as compared to pure curcumin. (21)

The phospholipids and different phytoconstituents (flavonoids) form a complex

between the poorly water soluble phytoconstituents and made stable complexes that

increase the bioavailability and reduces the side effects. This type of complex

formulation called the phytosomes. The phytosomes technology produces a little cell,

10

better able to transit from a hydrophilic environment into lipophilic environment of the

enterocyte cell membrane and from there into the cell, finally reaching the blood (22).

HO

H

H

H

NO

PO

O

O

H2C CH

O

CH2

O C

O

R1

C

O

R2+

OH

H

H

H

NO

PO

O

O

H2CCH

O

CH2

OC

O

R1

C

O

R2

O

HO

H

HO

HO

OH

HO O

O

HH

H

OP

OO

CH2

CHO

CH2

O

C

OR1

C

O

R2

O

O

HO

H

HO

HO

O

HO O

Cholesterol

Quercetin

Phosphatidylcholine

H

N

H

CHCl3Step-1

Step-2

Fig.1: Chemical reactions for formation of Cholesterol-Phosphatidylcholine-Quercetin complex

Cholesterol-Phosphatidylcholine-Quercetin complex

11

Phytosome are created when the standardized extract and active ingredients of an

herb are bound to the phospholipids on a molecular level. Phytosome structures contain

the active ingredients of the herb surrounded by the phospholipids (For the improvement

of nutrient absorption and bioavailability). (23)

The phospholipids are readily compatible with the entire range of vitamins, minerals,

metabolites, and herbal preparations currently consumed as the dietary phospholipids and

omega-3 fatty acid works in functional synergy in cell membranes. The phospholipids

mainly employed to make phytosomes is phosphatidylcholine, derived from soybean

(glycine max). Phosphatidylcholine is a bifunctional compound miscible both in water

and in oil environments, and is well absorbed when taken by mouth. (24)

1.1. Difference between phytosome and liposomes

In liposome the active principle is dissolved in the medium contained in the cavity or in

the layers of the membrane, whereas in the phytosome it is an integral part of the

membrane, being the molecules anchored through chemical bonds to the polar head of the

phospholipids.(25, 26)

1.2. Advantages of Phytosomes (27, 33, 34, 35)

1. It enhances the absorption of lipid insoluble polar phytoconstituents through oral

as well as topical route showing better bioavailability, hence significantly greater

therapeutic benefit.

2. As the absorption of active constituents is improved, its dose requirement is also

reduced and phosphatidylcholine used in preparation of phytosomes, beside acting

as a carrier, it also acts as a hepatoprotective agent, hence giving the synergistic

effect when hepatoprotective substances are employed.

3. Chemical bonds are formed between phosphatidylcholine molecule and

phytoconstituent, so the phytosomes show better stability profile.

4. Phytosome process produces a little cell, which is protect the herbal extract from

destruction by digestive secretion and gut bacteria and assured delivery to the

tissues.

12

5. Entrapment efficiency is high and more over predetermined because drug itself in

conjugation with lipids is forming vesicles so no problem of drug entrapment.

6. Phytosomes show better stability profile because chemical bonds are formed

between phosphatidylcholine molecules and phytoconstituents.

Phosphatidylcholine used in the phytosomes process besides acting as a carrier

also nourishes the skin, because it is essential part of cell membrane.

1.3. Methods of preparation

Following steps are involved in the preparation of phytosomes (31)

Passive Loading Techniques

1. Mechanical Dispersion Methods

2. Lipid film hydration by hand shaking non-hand shaking or freeze drying

3. Micro-emulsification

4. Sonication

5. Solvent evaporation method

1.4. Approaches to improve bioavailability

In case of poorly absorbed natural derived ingredients, various strategies are being

followed in the nutraceutical sector to achieve this goal. The first one might also seem to the

medicinal chemistry approach: by the chemical derivatization of the chemical product, the aim is

to obtain compounds showing an improved bioavailability. This approach, however, generates a

number of chemical analogues that need to be appropriately screened. An alternative strategy

that is also being pursued is the combination of the active molecules with other compounds as

adjuvants promoting the active molecule‟s absorption.

In this approach involves extensive formulation research of structures capable of both

stabilizing natural molecules and promoting their intestinal absorption. The formulative research

comprises the formation of liposomes, micelles, nanoparticles, nanoemulsions, microsphere or

other complexes. Many other approaches have been developed to improve the oral

bioavailability, such as inclusion by solubility and bioavailability enhancers, structural

modification and Entrapement with the Lipophilic Carriers such as phospholipids. (32, 33)

Well known example of the above mentioned approach is that the fruit of milk thistle

plant (Silybum Marianum, family Asteraceae) contains flavonoid that are proven liver

13

protectants. The standardized extract known as silymarin contains three flavonoid of the flavonol

subclass. Silybin predominates, followed by silydianin and silychristin. Although silybin is the

most potent of the flavonoid in milk thistle, similar to other flavonoid it is not well-absorbed.

Silybin-phosphatidylcholine complexed as a phytosome provides significant liver protection and

enhanced bioavailability over conventional silymarin. (34) These are main reasons for the growing

interest in spherical unilamellar vesicles: as they have become potentially important for drug

encapsulation of chemical or herbal origin, the latter use in their ability to enclose water-soluble

therapeutic molecules such as drugs and enzymes and to direct these compounds to specific drug

absorption. (35)

1.5. Merits of phytosomes over conventional dosage forms

a). There is a dramatic enhancement of the bioavailability of botanical extract due to their

complexation with phospholipids and improved absorption. They permeate the non-lipophilic

botanical extract to allow better absorption from the intestinal lumen, which is otherwise not

possible with other conventional dosage form, example: powder, tablet and solutions etc.(36)

b). They have been used to deliver liver-protecting flavonoid because they can be made easily

bioavailable by phytosomes. In addition to this phosphatidylcholine is also hepatoprotective and

so provide a synergistic effect for liver protection. This technology offers cost-effective delivery

of phytoconstituents and synergistic benefits. They can also be used for enhanced permeation of

drug through skin for transdermal and dermal delivery. (37)

c). Phosphatidylcholine, an essential part of the membrane used in phytosome technology, act

as a carrier and also nourishes the skin. There is no problem with drug entrapment during

formulation preparation. Also, the entrapment efficiency is high and more over predetermined,

because the drug itself forms vesicle after conjugation with lipid. They offer a better stability

profile because chemical bonds are formed between the phosphatidylcholine molecule and

phytoconstituents. (37)

d). The phytosomal system is passive, non-invasive and can is suitable for immediate

commercialization. The dose requirement is reduced due to improved absorption of the main

constituent. Relatively simple to manufacture with no complicated technical investment required

for the production of phytosomes. (38)

14

e). Phytosomes are also superior to liposomes in skin care products while the liposomes are an

aggregate of many phospholipids molecules that can enclose other photoactive molecules but

without specifically bonding to them. (38)

f). However, in the case of phytosomes products, numerous studies have proved that they are

markedly better absorbed and have substantially greater clinical efficacy. Companies have

successfully applied this technology to a number of standardized flavonoid preparations. (38)

Table 1.5: Different Type of Somes with Their advantages & disadvantages

Vesicular drug

delivery system

Method of

preparation

Advantages Disadvantages

Liposomes, which

encapsulate water and

lipid-soluble

pharmacologically and

cosmetically active

components.

Physical dispersion

involving hand

shaking and non-

hand shaking

methods

solvent dispersion

involving ethanol

injection, ether

injection, double

emulsion vesicle

method, reverse

phase evaporation

vesicle method, and

stable plurilamellar

vesicle methods

detergent

solubilization

Provide selective

passive targeting to

tumor tissues,

increased efficacy &

therapeutic index,

reduction in toxicity,

site avoidant effect,

improved

pharmacokinetic

effects

Degradation by

hydrolysis,

Oxidation,

Sedimentation,

Leaching of drug;

and aggregation

or fusion during

storage.

Phytosomes are

standardized extract or

purified fractions

Physical dispersion

involving hand

shaking and non-

It enhances the

absorption of lipid

insoluble polar

Phytoconstituent

is rapidly

eliminated from

15

complexed with

phospholipids for a

better bioavailability

and enhanced activities.

handshakes methods.

phytoconstituents,

Dose requirement is

reduced, Better

stability profile,

protect herbal extract

from destruction in

gastric fluids, and

assured delivery to

the tissues,

entrapment efficiency

is high.

herbosomes.

Ethosomes are

noninvasive delivery

carriers that enable

drugs to reach the deep

skin layers and /or the

systemic circulation.

Ethosomes contain

phospholipids, alcohol

(ethanol and isopropyl

alcohol) in relatively

high concentration and

water.

Ethosomes can be

prepared from

soyabean

phosphatidylcholine,

ethanol, drug should

be dissolved in

ethanol. Water added

in small quantity and

the preparation

mixed by mechanical

stirring under

controlled

conditions6

Ethosomes are

enhanced permeation

of drug through skin

for transdermal and

dermal delivery,

Peptides, proteins

molecules are better

transferred,

Ethosomes are safe

and approved for

pharmaceutical use,

High patient

compliance,

Simple to

manufacture.

Drugs that require

high blood levels

cannot be

administered,

Ethosomal

administration

cannot be given

in the form of

bolus type drug

input,

Molecular size

should be

reasonable for its

passage through

skin,

Adhesives may

not adhere to all

type of skin types,

Uncomfortable to

wear,

16

May not be

economical,

Skin irritation due

to excipients.

Pharmacosomes are the

colloidal dispersions of

drugs covalently bound

to lipids and may exist

as ultrafine vesicular,

micellar, or hexagonal

aggregates, depending

on the chemical

structure of the drug-

lipid complex.

The idea for

development of the

vesicular

pharmacosomes is

based on surface and

bulk interactions of

lipids with drug. Any

drug possessing an

active hydrogen

atom(-CCOH,-OH,-

NH2) can be

esterified to the lipid,

with or without

spacer chain that

strongly result in an

amphiphilic

compound, which

will facilitate

membrane, tissue, or

cell wall transfer, in

the organism.

Entrapment

efficiency high,

simple process, drug

covalently linked so

no leakage problems,

no entrapment

problem.

On storage,

undergo fusion

and aggregation,

as well chemical

hydrolysis.

Niosomes are nonionic

surfactants vesicles and,

as liposomes, are

bilayered.

Niosomes formulated

by:

lipid layer hydration

method reverse

phase evaporation

method

Patient compliance,

accommodate drug

molecule with a wide

range of solubilities,

characteristics of

vesicles formulation

Aggregation,

fusion, leeaching

or hydrolysis of

entrapped drugs,

thus limiting the

self-life of

17

transmembrane pH

gradient uptake

process(remote

loading)

hand shaking

ether injection

Sonication

are variable and

controllable,

osmotically active and

stable,

Biodegradable &

biocompatible, non

immunogenic.

niosomes

dispersion.

Niosomes

preparation is

time-consuming,

requires

specialized

equipment.

Transferosomes are

specially optimized,

ultradeformable lipid

supramolecular

aggregates, which are

able to penetrate the

mammalian, skin intact

& consist of at least one

inner aqueous

compartment, which is

surrounded by a lipid

bilayer with specially

tailored properties, due

to the incorporation of

“edge activatore” into

the vesicular membrane.

Transferosomes are

prepared in two

steps.first, a thin

film, comprising

phospholipid and

surfactant is

prepared, hydrated

with buffer (pH6.5)

by rotation and

desired size by

sonication.in second

step, sonicated

vesicles are

homogenized by

extrusion through a

polycarbonate

membrane.

Transferosomes are

good candidates for

the non-invasive of

delivery of small,

medium,and large

sized drugs. Multiliter

quantitiesof sterile,

well-defined

transferosomes

containing drug can

be prepared relatively

easily

They are

completely

degraded in the

GI tract, and

when used in a

degradation

preventing

formulation, their

uptake in the gut

becomes

problematic and

extremely

insufficient.

18

Table 1.6: Liposomal herbal formulations and their applications

Name of Bioactive Component/Plants Applications References

Essential oil from Atractylodes macrocephala koidz Increase in solubility &

Bioavailability

39

Essential oil of O. dictamnus Increase in activity 39

Extract of tripterygrium wilfordi Reduction in side effects 40

Quercetin Increase in bioavailability 41

Silymarin extract Increase in hepatoprotective

activity

42

Essential oil of Artemisia arborescens L. Increase in stability 43

Texans Decrease in toxicity 44

Table 1.7: Phytosomal Herbal Products

Phytosomes Source Uses

Silybin Phytosomes Silybin flavonoids from Silybrium

marianum

Hepatoprotective, antioxidant for

liver

Ginkgo Phytosomes Ginkgo flavonoids from Ginkgo

biloba Protects brain and vascular linings

Ginseng Phytosomes Ginsenosides from Panax ginseng Nutraceutical and immunomodulatory

Green tea Phytosomes Epigallocatechin from Thea

sinensis Anti-cancer, Nutraceutical

Grape Seed Phytosomes Procyanidins from Vitis vinifera Nutraceutical, systemic antioxidant,

cardio protective

Olive oil phytosomes Polyphenols from olea europaea

oil Antioxidant, anti-inflammatory

19

1.6. REVIEW OF LITERATURE

1. A. Choubey (2011): reviewed enhancement of bioavailability of some orally administered

botanical extract which is erratic and poor due to limited gastro-intestinal absorption.

Bioavailability can be improved by using new delivery systems which can enhance the rate and

the extend of solubilization into intestinal fluids and capacity to cross biomembranes and current

trends in phytosomes drug delivery and it was concluded that phytomedicines, complex chemical

mixtures prepared from plants, have been used in medicine. phytosomes are advanced forms of

herbal product that are better absorbed, utilized, and as a result produce better results than

conventional herbal extracts.(45)

2. T. P. Raju et al (2011): reported that advanced biochemical and pre-chemical studies have

proved the potential of plant flavonoid, polyphenolics and other hydrophilic natural compounds,

for the treatment of skin disorders, different types of carcinoma, anti-aging and many other areas

of the therapeutics and preventive medicine. The hydrophilic nature and unique chemical

structure of these compounds pose major challenge because of their poor bioavailability through

the skin or gut. The bioavailability can be improved by the use of novel delivery systems, which

can enhance the rate and the extend of drug solubilization and capacity to cross the lipid rich

biomembranes. At last concluded that phytosomes of enhanced bioavailability have improved

pharmacokinetic and pharmacological parameter, which in result can advantageously be used in

treatment of various acute diseases as more amount of action (liver, brain, heart, kidney etc.) at

similar or less dose as compared to the conventional plant extract. (46)

3. A. Rajendra et al (2011): Studied phytosomes technology which enhances the bioavailability

of herbal extract. In phytosomes technology the individual components of an herbal extract are

bound to phosphatidylcholine. Plant extract can be standardized accordingly and may be

formulated as phytosomes for systemic investigation for any improved potential use and give the

significant therapeutic or health promoting properties when compared with the conventional

plant extract. (47)

4. M. Sunitha et al (2010): Studied phytoconstituents despite having excellent bioactivity in

vitro, demonstrates less or no in vivo actions due to their poor lipid solubility or improper

molecular size or both, resulting in poor absorption and poor bioavailability. Lipid solubility and

molecular size are the major limiting factor for molecules to pass the biological membrane and to

be absorbed systematically following oral or topical administration and concluded that

20

phytosomal potential in cosmetics, as anti-inflammatory, in cardiovascular disorder, immune-

modulator, anti-cancer, anti-diabetic, etc.(48)

5. K. Prashant et al (2010): reviewed phytosome preparation by non-conventional method.

Absorption of phytosomes in gastro-intestinal tract is appreciably greater resulting in increased

plasma level than the individual component. Phytosomes form bridge between the conventional

delivery system and novel delivery system. (49)

6. S. Sharma et al (2010): reviewed phytosomes are novel drug delivery system containing

hydrophilic bioactive phytoconstituents of herbs surround and bound by phospholipids. This

technology has improved pharmacokinetics and pharmacological parameters of plant extract and

provided the good treatment of various diseases of human being. (50)

7. S. Pandey et al (2010): Demonstrated that most of the bioactive constituents of

phytomedicines are flavonoid and majority of the flavonoid are poorly bioavailable when taken

by mouth only a small fraction of a given dose reaches the blood and present the evidence that by

converting certain flavonoid nutrient to their phytosomes equivalent, Indene‟s proprietary

process increases their bioavailability by 2 xs to 6xs, and more and concluded phytosome

preparation was found to have at least 10 potential action mechanisms, all operative at

concentrations that can be realistically attained in humans.(51)

8. P. G. Sindhumol et al (2010): Reviewed the effectiveness of any herbal medication is

dependent on the delivery of effective level of the therapeutically active compound. But a severe

limitation exists in their in their bioavailability when administered orally or by topical

applications. Phytosomes are recently introduced herbal formulation that are better absorbed and

as a result produced better bioavailability and actions than the conventional phytomolecules or

botanical extracts and concluded that phytosomes are novel compounds comprising of lipophilic

complexes of components of various plants like Silymarin Marianum, Ginkgo Biloba, ginseng

etc. with phospholipids, preparation of phytosomes is usually carried out by non-conventional

method, absorption of phytosome in gastro intestinal tract is appreciably greater resulting plasma

level than the individual component.(52)

9. K. R. Vinod et al (2010): Reviewed phytosomes, often known as herbosomes are recently

introduced in herbal formulation that exhibits better pharmacokinetics and pharmacodynamics

profile than conventional botanical extracts. They are produced by a process whereby the

standardized plant extract or its constituents are bound to phospholipids, mainly

21

phosphatidylcholine producing a lipid compatible molecular complex, the major molecular

building block of cell membrane and a compound miscible in both water and in oil or lipid

environments. This technique makes phytosomes unique in character as potential, concluded

phytosomes are advanced form of herbal extract that are better absorbed which result better than

conventional herbal extract.(53)

10. J. Naga Sowjanya et al (2010): Studied the most of the prevailing diseases and nutritional

disorders are treated with natural medicines. The novel formulation for natural components,

“phytosomes” which are better absorbed, utilized and as a result produced better results than

conventional herbal extracts owing to the presence of phosphatidylcholine which likely pushes

the phytoconstituent through the intestinal epithelial cell outer membrane, subsequently

accessing the bloodstream an can be advantageously be used in the treatment of the acute and

chronic liver disease of toxic metabolic or infective origin or of degenerative nature and

concluded phytosomes exist as bridge between the conventional delivery system and novel

delivery system.(54)

11. Nilesh Jain et al (2010): Reviewed that the term phytosomes which is advanced forms of

herbal formulations which contains the bioactive phytoconstituents of herb extract surrounds and

bound by a lipid. Most of the bioactive constituents are water-soluble compounds like flavonoid,

glycosides; terpenoid in which flavonoids are a major class of bioactive compounds possesses

broad therapeutic activities. Because of water soluble herbal extract and lipophilic outer layer

phytosomes shows better absorption and as a result produce better bioavailability and actions

than the conventional herbal extracts containing dosage form. These extract can be standardized

accordingly and may be formulated as phytosomes for systemic investigation foe any improved

potential to be used rationally and thus different therapeutic purposes like cardiovascular, anti-

inflammatory, immunomodulatory, anticancer, anti-diabetic etc or for prophylactic and health

purposes as nutraceutical.(55)

12. K. Kaku 2010: Reviewed that impaired insulin secretion and increased insulin resistance, the

main pathophysiological features of type 2 diabetes, jointly contribute to the development of this

disease. Recently, it has become widely recognized that the functional pancreatic cell mass

decreases over time and type 2 diabetes is a progressive disease. Studies suggest the possibility

that the Japanese may have many genes susceptible to diabetes including thrifty genes. Various

environmental factors, added to these genetic factors, are considered responsible for the onset of

22

disease, and the number of patients is increasing rapidly reflecting recent lifestyle changes.

Impaired insulin secretion is characterized by lowered glucose responsiveness. In particular, the

decrease in postprandial-phase secretion is an essential pathophysiological condition.

Glucolipotoxicity, if left untreated, results in the decrease in the functional pancreatic cell mass.

The goal of diabetes treatment is to secure a quality of life (QOL) and lifespan comparable to

those of healthy people, and a prerequisite for this is the prevention of onset and progression of

vascular complications. The need for earlier initiation of proactive intervention must be

emphasized, as well as the importance of comprehensive (blood sugar, blood pressure, and

lipids) intervention in attaining this goal. (56)

13. Jagruti patel et al (2009): Studies the advanced biochemical and pre-clinical studies have

proved the potential of plant flavonoid and other hydrophilic natural compounds for the

treatment of skin disorders, different types of carcinoma, anti-aging and many other areas of

therapeutics and preventive medicine. The hydrophilic nature unique chemical structure of these

compounds poses major challenge because of their poor bioavailability through the skin or gut.

(57)

14. Sanjib Bhattacharya et al (2009): Studied the lipid solubility and molecular size are the

major limiting factors for molecules to pass the biological membranes to be absorbed

systematically following oral or topical administration. Several plant extract and

phytoconstituents, despite having excellent bio activity in vitro demonstrates less or no in vivo

actions due to their poor lipid solubility or molecular size or both, resulting poor absorption and

poor bioavailability. Phytosomes are advanced form of herbal formulations that are better

absorbed and as a result produce better bioavailability and actions than the conventional herbal

extract. (58)

15. N. S. Chauhan et al (2009): Reviewed the potential uses of large number of herbal drugs are

limited due to their poor absorption and poor bioavailability after oral administration. The

bioavailability can be improved by formulating appropriate drug delivery system, which can

enhance the rate and extend of drug absorption across the lipoid biomembranes. Phospholipids

based drug delivery system have been found promising for better and effective delivery of drug

and providing much appropriate systematic drug delivery. The phospholipids molecular structure

includes a water-soluble head and two fat- soluble tails, because of this dual solubility, the

phospholipids acts as an effective emulsifier, which is also one of the chief components of the

23

membranes in our cells and concluded that phytosomes are better absorbed which result better

than conventional herbal extract. (59)

16. S. Bhattacharya et al (2009): Reviewed the several plant extracts and phytoconstituents,

despite having excellent bio-activity in vitro demonstrate less or no in vivo actions due to their

poor absorption and poor bioavailability. Phytosomes exhibit better pharmacokinetic and

pharmacodynamics profile than conventional herbal extracts and concluded in recent times the

emerging technology of drug delivery and drug targeting is also being applied to

phytopharmaceuticals. Botanicals have enormous therapeutic potential which should be explored

through some value added drug delivery systems. (60)

17. D. M. Brahmankar et al (2006): Mentioned in case of poorly absorbed natural derived

ingredients, various strategies are being followed in the nutraceutical sector to achieve this goal.

The first one might also seem to the medicinal chemistry approach: by the chemical

derivatization of the chemical product, the aim is to obtain compounds showing an improved

bioavailability. This approach, however, generates a number of chemical analogues that need to

be appropriately screened. An alternative strategy that is also being pursued is the combination

of the active molecules with other compounds as adjuvants promoting the active molecule‟s

absorption. A third approach involves extensive formulation research of structures capable of

both stabilizing natural molecules and promoting their intestinal absorption. The formulative

research comprises the formation of liposomes, micelles, nanoparticles, nanoemulsions,

microsphere or other complexes. Many other approaches have been developed to improve the

oral bioavailability, such as inclusion by solubility and bioavailability enhancers, structural

modification. (61)

18. K. Parris et al (2005): Reviewed bioavailability enhancement of water-soluble flavonoid

molecules can be converted into lipid-compatible molecular complexes, known as phytosomes.

The fruit of the milk thistle plant contains flavonoids that are proven liver protectants. Silybin-

phosphatidylcholine complexed as a phytosome provides significant liver protection and

enhanced bioavailability over conventional silymarin and concluded that silybin-PC complexed

as a phytosomes provides significant liver protection and enhanced bioavailability over

conventional silymarin when taken orally. (62)

19. A. D. L. Maza et al (1998): Studied the interaction of the amphoteric surfactant

dodecylbetaine with phosphatidylcholine (PC) liposomes was studied by means of transmission

24

electron microscopy (TEM) and changes in the mean particle size. A good correlation was found

between the TEM diameter of the particles and the mean hydrodynamic diameter (HD)

determined by QELS. (63)

25

1.7. OBJECTIVE OF RESEARCH:

Many advances have taken place in the field of anti-diabetic drugs and a large

number of drug development techniques have been used to prepare herbal anti-diabetic

formulations because of the adverse effects from the allopathic drugs on the long therapy.

Generally, herbal anti-diabetic plant or plant extracts are having two major limiting

factors such as lipid solubility and molecular size to pass the biological membrane, both

factor affect the bioavailability of plant extract, when given by oral or topical route. Since

several plant extracts and phytoconstituents, despite having excellent bioactivity in vitro

have less or no in vivo actions due to their poor lipid solubility or improper molecular

size or both, resulting poor absorption and poor bioavailability. Some constituents of the

multi-constituent plant extract are destroyed in gastric environment when taken orally.

Various phytoconstituents (flavonoids) are poorly water soluble in nature, and

need to large dose size to obtained desired pharmacological action. The phospholipids &

plant constituents (flavonoids) form a complex and made stable complexes that increase

the bioavailability and reduces the side effects associated with dose size. This type of

complex product known as phytosomes.

There are many types of solubility enhancement techniques, but out of them

physical modification is the most common, i.e. solid dispersions, microemulsions,

eutectic mixtures and complexation etc. complexation is the technique in which two or

more molecules associated in form of bonded/non-bonded entity with a well-defined

stoichiometry. Various complexing agents such as EDTA, cyclodextrins and polymer

have been used for the complexation.

The phytosomes enhance the bioavailability by modifying the solubility of plant

extracts, in which technique phospholipid form the complex with phyto constituents.

Phytosomes are advanced forms of herbal formulations that are better absorbed, produce

better bioavailability than conventional herbal extract. The phospholipids are readily

compatible with the entire range of vitamins, minerals, metabolites, and herbal

26

preparations currently consumed as the dietary phospholipids and omega-3 fatty acid

works in functional synergy in cell membranes.

Cordia Dichotoma / Carissa Carandas have been reported for various

pharmacological activities but detailed studies regarding antidiabetic activity has not been

reported. The work was taken up to investigate the anti-diabetic activity of various

solvent fractions and components from the seeds which have been mentioned in

traditional system for use in tridosha and in diabetes. So the aim of research to establish

the quantitative correlation between the photochemical, pharmacological profile and also

development of the novel herbal formulation for diabetes mellitus patients.

Pharmaceutical scope: phytosomes have following advantages.

It enhances the absorption of lipid insoluble polar phytoconstituents.

Dose requirement is also reduced.

Phosphatidylcholine acting as a carrier also acts as a hepatoprotective hence

giving the synergistic effect when given with hepatoprotective formulations.

Chemical bonds are formed between phosphatidylcholine molecule and

phytoconstituent, so the phytosomes show better stability profile.

Protect the herbal extract from destruction by digestive secretion and gut bacteria.

Entrapment efficiency is high and more over predetermined because drug itself in

conjugation with lipids is forming vesicles so no problem of drug entrapment.

27

PLAN OF WORK

1. COLLECTION OF PLANT MATERIAL

2. PHARMACOGNOSTIC STUDIES

A. Macroscopically evaluation

B. Microscopically evaluation

C. Physiochemical studies

D. Determination of extractive value

3. PRELIMINARY PHYTOCHEMICAL SCREENING

4. CHROMATOGRAPHY ANALYSIS

A. HPTLC

5. PHARMACOLOGICAL SCREENING

6. PREFORMULATION STUDIES

A. Spectroscopy methods

B. Phytoconstituents –lipid interaction study

C. Standard curve of marker compound

7. FORMULATION DEVELOPMENT

A. Selection of lipid

B. Plant extract ratio

8. EVALUATION AND CHARACTRIZATION

A. Quantification of CDL CCL

B. Size analysis of Phytosomes

C. TEM

D. Separation of entrapped and unentrapped plant extract

E. Zeta Potential

F. Invitro Release studies

9. STABILITY STUDIES

-Short term & Long term

28

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33

CARISSA CARANDAS

CORDIA DICHOTOMA