Connect with us

Pharma

Rice Ferment Filtrate in Cosmetics

Yesenia Harris

Published

on

About Author

Vinay Kumar Singh.

Head-Formulation

Kumar Organic Products Research Centre Pvt. Ltd., Bengaluru

Rice water is nothing but cloudy-milky water that you throw away after soaking the rice for a few hours or boiling them. You would be surprised to know that rice water contains a lot of minerals and vitamins like amino acids and vitamin B6, which might benefit your skin and hair by increasing shininess, enhancing softness, strengthening the hair while protecting against the various damages to your skin & health goes through.

Rice is part of India’s beauty legacy, which contains ferulic acid, a powerful antioxidant, and allantoins, anti-inflammatory properties. So, whether you eat rice or not, use it in your face pack for glowing skin. The starch in rice water soothes the skin in conditions like eczema.

Rice water has been used in Asian beauty rituals for centuries to achieve glowing and smooth skin. Using rice water for the skin is a natural home remedy in treating and preventing acne, removing acne scars, and getting rid of hyperpigmentation.

Rice water contains amino acids, vitamins B and E, and minerals that promote cell growth, stimulate blood flow and keep the skin smooth, youthful, and bright in chilly weather conditions. It’s a beauty booster used by more and more people worldwide.

Rice is, of course, delicious as one of the vital culinary ingredients worldwide, so we all know how to make it, but when we are using it for beauty purposes, it’s a different process, very simple, by the way.

A Chinese Village in Huangluo was named as the ‘Long Haired Village’. This is because the Yao women out there have the most beautiful, shiny and long hair. And guess the length of their hair? The majority of the women have hair longer than 1.4 meters, and they even made a place in the Guinness Book of World Records as the “world’s longest hair village”.

Yao women have a belief that long hair signifies prosperity, good fortune, longevity, etc. Thus their long hair is undoubtedly their prized possession! Also, they do not have grey hair till their late 80s.

The Yao women believe that the fermented rice water, which they use to cleanse their hair, is what helps to keeps their hair long, dark and clean.

Furthermore, female rice farmers in China, Japan, and other Southeast Asian countries used to bathe and wash, in the water used for cleaning rice. Rice water is the secret recipe of flawless-skinned Japanese geishas for centuries.

In Korean cuisine, rice water has been used for a long time to cook dishes like stews. They also use rice water for braising fish or to wash it, which further takes away all the smell of the fish. Korean people also use rice water for deodorizing their plastic containers. You will be surprised to know that it can also be used for cleaning dishes and as a natural fertilizer for plants as it’s loaded with minerals and vitamins.
Benefits of Rice water for hair and skin

1. Promotes hair growth

The fascinating length of the Yao women proves that rice water inevitably has a significant role in the hair growth. This magical potion is rich in Amino Acids, Vitamin B, Vitamin C and Vitamin E, which encourages hair growth to a great extent. The Amino Acids are an excellent ingredient for hair growth. It boosts

protein and keratin production, which are the basic necessities for your hair. Vitamin B is significant to maintain your hair’s strength, and Vitamin C keeps the sebum production in check to an optimum mark. Vitamin E prevents hair loss and keeps your hair hydrated so that it doesn’t feel dry or dull.

2. Prevents Split Ends

The problem of Split ends is pretty common, and this is a real nuisance. The amount of effort and time invested in making your hair long gets wasted completely when you chop off your hair length to eliminate the split ends. This is simply heart breaking. Rice water is an excellent remedy to prevent split end.

3. Cures Dandruff

There is simply no individual on earth who hasn’t dealt with the struggle of dandruff. Rice water has been proven to help with dandruff by fighting the fungi, which causes dandruff and eliminates those dry white flakes to a large extent. It also helps maintain the pH level of your hair.

4. Gives damage protection

Styling tools are common in our everyday life. Perfect straight hair is one of the ongoing trends right now. Investing in a damage protectant or protectant serum is essential but can be costly. The carbohydrate present in the rice water is a great damage protectant. It can be used before applying any styling tools. It reduces the surface friction and also increases the elasticity of your hair. Thus, making it less prone to damage by those heating tools.

5. Has Inositol, a carbohydrate to help

A recent research showed that rice water exhibits hair care properties, such as decreasing surface friction and improving hair elasticity. Rice water has inositol, a carbohydrate that can repair damaged hair, as well as protect it from further damage. Specialized imaging technique shows that inositol stays inside the hair even after rinsing, offering continuing hair protection and beautifying effects. These remarkable benefits explain the 6 feet, healthy, long and beautiful hair of the Yao women.

6. Skin benefits.

Due to its cooling and soothing effects on skin, it is often prescribed as an effective ointment to cool off inflamed skin. Rice water also has moisturizing, antioxidant, and healing properties that help to improve circulation, prevent or fade age related spots, and ease inflammation to give you a healthy, better moisturized and clear skin. As long as it’s left on the skin, rice water is even believed to offer mild protection from the sun.

Rice water illuminates, firms, and tightens skin to create a porcelain finish. It reduces pore size, leaving a powdery, soft feeling behind. Rice water is also good for acne since it reduces redness and blemishes.

Black rice is indigenous to Northeast India, and is extensively grown in Odisha, West Bengal and Jharkhand. Commonly eaten in Manipur, the dish is called chak-hao, which translates to rice (chak) which is delicious (ahaoba). It’s a medium-grain rice that originated in China and other parts of Asia some 10,000 years ago.

Varieties of black rice is grown everywhere from Thailand and Indonesia to China, Japan, and India. This rice also grows in countries all across the world. Some varieties are short-grained, and some are long-grained. Long-grained rice is slightly healthier since it has less starch, giving it a lower glycemic index.

Kavuni Arisi (Black Rice): Almost unique to the Chettinad region in Tamil Nadu and evidence of the Chettiar community’s strong links with Burma, black rice is prevalent across South East Asian cultures and is a rich source of Iron and Vitamin E. The bran hull of black rice has one of the highest levels of anthocyanin antioxidants found in any food (even more than blueberries).

Black rice has a spectacularly high level of anthocyanin, which might be why it’s revered in, for example, China as an extremely healthy food as they help to protect your cells against damage. These pigments are also known for reducing inflammation and helping reduce the risk of cardiovascular conditions. Eating black rice can give you a worthwhile boost on your heart health and overall fitness.

It also is fairly tricky to grow, at least in terms of yield, which is often as low as 10 percent of other rice varieties. It’s occasionally called “emperor’s rice” or “forbidden rice” for that very reason but oral history suggests that only the richest of the rich were able to afford the delicacy. Chinese emperor to ensure his health and longevity, had forbidden it to all others. In fact, it has more fiber and protein than red, brown, or white rice.

Black rice also provides these health benefits :

Improved Eye Health: Along with protective anthocyanins, black rice contains high amounts of lutein and zeaxanthin, two carotenoids known for their role in supporting eye health. These antioxidants help to protect the cells in your eyes and to reduce the effects of ultraviolet (UV) radiation.

Diabetes Management: Flavonoids like anthocyanins also influence blood sugar levels and diabetes management. Phytochemicals have a positive influence on our body, improving insulin sensitivity. They also help by reducing sugar digestion in your small intestine, which lowers sugar levels in your blood.

Nutrition : Black rice is rich in amino acids, fatty acids, antioxidants, flavonoids, anthocyanins, and other phenolic compounds. There are 18 amino acids in black rice, with a mix of essential and non-essential types. Amino acids are crucial to many of your body’s functions, from helping repair skin and tissues to improving your energy levels and digestion.

At KOPL Biotech Centre, we explored The SCIENCE behind Rice Water. We believe in innovation so we have launched Kopryza- A black rice ferment filtrate.

The pH level of rice water is slightly harsh on our hair. However, Fermentation process reduces the pH level of rice water and equal to the pH level of the Hair, and nourishes it better due to the effective penetration of vitamins into the scalp and hair structure. Its purported benefits include “promotes hair growth, protects and repairs hair, increases shine and smoothness, detangles the hair, reducing hair fall and makes hair stronger”

Fermented rice water contains “Vitamins, proteins, minerals, keratin and antioxidants”. Once fermented, rice water nutrition is magnified, it is found to produce additional nutrients such as niacin and biotin, which are ingredients in most of the hair growth supplements. Fermented rice water has a natural carbohydrate called Inositol. Inositol penetrates and lingers in the hair shaft to provide repair and lasting protection.

Among the various rice varieties researched at our Biotech Centre, Black Rice was found to possess significantly higher levels of different carotenoids like ?-carotene, lutein and phenolics. It is rich in 18 amino acids, minerals and vitamins like Copper, Iron, Zinc. It is also reputed for having high anthocyanin content and hence, high antioxidant activity, which protects, the skin and hair from damages caused by free radicals.

KOPRYZA is the new hair care ingredient developed at KOPL Biotech Centre where all the benefits of Karuppu, Kavuni Rice are scientifically captured for the benefit of the hair & skin. They are highly recommended for all kinds of shampoos and conditioners and for all types of hair. The best thing is that the goodness imparted onto the hair stays even after multiple washes. The original goodness of black rice water further gets augmented by fermentation.

Inositol content of Kopryza was estimated by HPLC method as 48.45mg Inositol /100g

Bioactives(Peptide) in Kopryza: Oligopeptide is a small molecule of protein, which is also known as a peptide. Oligopeptide content of Kopryza is found to be 5.81 g/L. The molecular weight of this peptide is found to be 10 KDa.

Keeping in mind, the attribute it offers, we have developed many formula. One such formulation of 2 in 1 Conditioning Shampoo, as described below which will help repair hair damage. It will wash away hair woes with the natural goodness of Fermented Black Rice Water and put a full stop to deteriorating hair quality.It improves hair texture by adding elasticity, increasing volume, and giving it a healthier appearance, while also shielding it from further damage being rich in vitamins and minerals.

FORMULATION

2 in 1 Conditioning Shampoo

This Shampoo tends to clean the hair by removing dead cells and sebum from scalp. Kopryza present in the shampoo conditions, helps in detangling the hair & making them smooth and shiny.

Phase

Sr.No

Ingredient/INCI

Wt.%

Trade name /supplier

Uses

A

1

Aqua

QS to 100

In-house

Solvent

2

Acrylates/C10-30 Alkyl Acrylate Cross polymer

0.3

Lubrizol

Rheology modifier

3

Sodium Lauryl Ether Sulfate

45

Godrej

Surfactant

4

Guar Hydroxy Propyl Trimonium chloride (GHPTC)

0.3

Akzo nobel

Rheology modifier

5

Dimethicone

0.75

Dow corning

Conditioning & Shining agent

6

Disodium EDTA

0.1

S.D.Fine

Chelating agent

7

Cocoamidopropyl Betaine

6

Delta Green

Surfactant

B

8

Cetyl Alcohol

0.6

S D Fine

Emulsifier

9

Cocomonoethanol amine

2

Otto Chemicals

Foaming agent

10

Ethylene Glycol Distearate

0.3

Croda

Pearlizing agents

C

11

Polyquaternium 44

1

Delta Green Science

Conditioning agent

12

Rice ferment Filtrate

2

Kopryza/ KRC

Conditioning agent

13

D Panthenol

0.6

Delta Green Science

Conditioning agent

D

14

Green Infusion

0.5

Ultra International

Perfume

15

Triethyl citrate, Benzoic acid, Caprylyl glycol

1

KOPTBC/ KRC

Preservative

E

16

pH adjuster (18% NaOH)

~0.5*

S.D Fine

pH Adjuster

*pH range- 6.0-7.0

Procedure :

1. Weigh phase A & phase B ingredients separately and heat to 70-80?C.

2. Add phase B at 75?C to Phase A at 75?C slowly under stirring, maintain temperature of 75?C for 5-10mints.

3. Add phase C to phase AB at 45?C.

4. Add phase D at Room temperature (RT) to phase ABC at 35-40?C under mixing. Mix it well and allow to cool to RT.

5. Adjust pH with phase E and fill the product in suitable container.

Product Specification :

Parameter

Specification

Results

Appearance

White/off white Viscous liquid

Off White viscous liquid

Odour

Characteristic

Characteristic

pH

4.0-9.0

6.01

Viscosity*

Min.3000cps

4500cps

*Note: Viscosity determined by using Brookfield viscometer, Model LVDV II+PRO,

Spindle no: S6.3 @ 50RPM, 26?C

Ref : FD/AT/10/023A

Stability: Passes 12weeks at 45?C and passes 3 freeze/thaw cycles

Conclusion: Product was observed to be stable without any colour change or separation. Thus it was found to be stable.

KOPL Products :

Kopryza : Rice ferment filtrate Obtained by fermentation of Black Kavuni, Rice a rare variety of rice from south India using Bacillus bacteria, Suitable in skin care & hair care products

KOPTBC : This is a preservative blend of Caprylyl glycol, Triethyl citrate, Benzoic acid which is used in skin care, hair care Color cosmetics products.

Directions for Use :

Wet hair thoroughly with warm water. Take shampoo in your hand and rub it to generate foam. Apply on your scalp and hair length. Massage the shampoo into your scalp for 2-3 minutes. Rinse your hair with plenty of water. Use conditioner for better result

Fermented Black Rice Filtrate has lingering effective conditioning effect for hair and soothing & moisturizing effect on skin, making it useful for Hair care, Skin care & Colour cosmetics alike and truly Rice water just might be Asia’s best kept beauty secret, which is also confirmed by The Health Science Journal.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT admin@pharmatutor.org

FIND OUT MORE ARTICLES AT OUR DATABASE

Read More

Original Post: pharmatutor.org

Pharma

Kokum Butter in Cosmetics

Yesenia Harris

Published

on

By

Vinay Kumar Singh.
Head-Formulation
Kumar Organic Products Research Centre Pvt. Ltd.,
Bengaluru
Email : formulation_krc@kopresearchcentre.net

Plant-derived oils and butters are among the most popular ingredients for a variety of personal care products including Cream, lotions, Lipstick, lip balm, Make- ups and hair treatments.

The mere mention of kokum brings to mind the small, round, red to purple-hued fruits, relished in curries for their sour flavour, besides being sipped on as a delightful sharbat and chilled juice. While the kokum fruits, scientifically termed Garcinia indica and native to the Western Ghats region of Maharashtra, Karnataka, Goa, are extensively used in cooking traditional Indian dishes, the flattened, black, pliable seeds within yield a rather beneficial substance too, an inherently oily element known as kokum butter and this plant-based matter derived from the kernels of kokum, a crop belonging to the same botanical family as mangosteen, confers astounding merits for glowing skin and silky hair. Kokum fruit is an inseparable part of Konkani cuisine.

Kokum butter is an oleaginous material isolated from the seeds of the kokum fruit. Each raw or ripe kokum fruit contains about 5 to 10 large black seeds, which are separated from the pulp. They are then wiped clean to remove all dirt and debris, squeezed under high pressure and subsequently processed as vegetable oil to obtain kokum butter, otherwise known as kokum oil.

Kokum butter possesses a firm texture and usually appears in shades of light grey, pale white, creamy yellow. It is in fact a hard edible butter at room temperature, used as a substitute for cocoa butter in preparing confectioneries, as well as for topical use on skin and hair. Nevertheless, owing to its ease of melting upon touching the skin and umpteen valuable phytochemical compounds, kokum butter is widely incorporated into commercial personal care products.

Kokum Butter is high in essential saturated fatty acids like Omega 6 and Omega 3 which prevent regularly skin damage by making Skin Healthier and Balancing Moisture in the skin.
Unrefined Kokum butter provides the skin with extreme hydration and moisturization. It consists of a high Anti-inflammatory property that helps to reduce inflamed skin and issues like allergies, infections, rashes, and irritation. Kokum Butter is also enriched with Antioxidants and Vitamin E which helps to immune the skin against free radicals and toxins.

The moisturizing agent in the Raw and Pure Kokum butter prevents the skin from dehydrating and promotes skin cell regeneration. It also combats the visible signs of aging by preventing issues like decoloration, fine lines, and wrinkles. Kokum butter not only Benefits Skin Health but can also boost the Health of the Immune System and cell functioning.

Whipped Kokum Butter is proven to be beneficial for hair by promoting Hair Growth. It locks the moisture deep into the skin and nourishes the itchy & dry scalp and also reduces Dandruff & Hair Fall. Organic and unrefined Kokum butter is an ideal ingredient for lip balms, lip gloss, lotions, moisturizers, and ointments because of the presence of rich emollients in it.

Kokum butter is Non-comedogenic so it does not clog pores or cause acne. You can also use whipped Kokum butter for eczema and acne scars. When it is applied on a regular basis the skin will automatically soften and restore its elasticity. The shelf life of Kokum butter is quite long as compared to any other body butter because of the high oxidative stability.
The advantages of kokum butter include :
o No scent. Kokum butter naturally has no scent. Cocoa, coconut, and shea butters are well known for their distinctive fragrances. Forperson sensitive it fragrance, kokum butter may be a better option.
o Easily absorbed. Unlike most other plant butters, kokum butter is remarkably light, absorbed quickly and easily, and not greasy.
o Doesn’t clog pores. U-nlike shea butter, kokum butter won’t clog your pores or cause acne.
o Very structurally stable. Kokum butter is one of the most structurally and chemically stable plant butters available. It works great as a natural emulsifier or hardening agent for homemade cosmetics.

Benefits of Kokum Butter for Skin and Hair
Kokum seeds contain vitamin E and many powerful antioxidants. These nutrients strengthen the immune system and cell function, and help reverse the damage caused by free radicals.1. Skin
It’s no surprise that kokum butter is the ingredient makeup artists often use. This ingredient is loved for its highly nourishing properties. It helps create a smooth texture on the skin. This omega-rich butter can be applied directly on lips, hands, knees, and elbows. Following are benefits Kokum butter has for skin.
o Improves skin cell regeneration
This ingredient is also known for regenerating skin cells. At the same time, it effectively reverses skin cell degeneration, and thus prevents damage even before it occurs. It is a natural emollient, and can thus go to the deepest layers of the epidermis. This helps to heal wounds and chapped skin.
o Reduces visible signs of aging
It is also believed that this buttery ingredient helps address and prevent multiple skin aging signs. These include hyperpigmentation, increased fragility, thinning of the skin, reduced elasticity, dehydration, and dark spots. It also helps create a protective moisture barrier against pollution and seasonal changes.
o Deeply Moisturizes dry skin
It is best known for its ability to be an intense moisturizing agent. It can be used to restore the skin’s moisture content, including your lips, feet, hands, etc. Unlike other kinds of butter used in skin care, kokum oil or butter is not sticky.
It is lightweight, gets easily absorbed, and leaves no signs of greasiness after application. This is why most skincare experts advise it for people with sensitive skin.
o Treats acne
This butter has a strong moisturizing ability and is considered non-comedogenic, which means it does not clog pores. It restores moisture content to dry or irritated skin.
o Reduces skin inflammation
It can help ward off signs of inflammation on the skin regardless of the cause. It also prevents the risk of future inflammation by safeguarding the skin against skin aggressors.
o Best for sensitive skin
Known for its healing and anti-inflammatory properties, this naturally-occurring emollient makes the best alternative for sensitive skin. If your skin is sensitive to most skin care products, this ingredient is gentler and easily tolerated by the skin. So, even people with sensitive skin can use kokum butter for skin lightening and brightening.

2. Hair
This nourishing butter is equally helpful in hair care too. Below are some of the most important ways in which this natural butter helps hair.
o Anti-dandruff solution
There are common hair care issues like itchy or flaky scalp, hair loss, and other concerns. Its anti-inflammatory properties help reduce infection and inflammation on the scalp.
It also keeps the scalp moisturized. This is crucial while preventing dandruff and seasonal dryness, and thus creating a healthy scalp environment.
o Natural hair conditioner
Kokum butter is saturated with omega-3 fatty acids and other vital moisturizing agents that give the hair the right amount of hydration. It moisturizes the scalp and prevents oxidative stress in the hair cuticles.
o Stimulates hair growth
This butter is popularly known to stimulate hair growth. This natural plant butter is a perfect remedy for thick and long hair. It is rich in antioxidants, fatty acids, and lauric acid content. The fatty acids nourish the scalp and form a protective barrier against environmental or seasonal changes.

Read More

Source: pharmatutor.org

Continue Reading

Pharma

Lack of Medicines in Greece and the Solution of Generics

Yesenia Harris

Published

on

By

The problem of drug shortages is taking on alarming proportions worldwide. In Greece at the moment, antibiotics, antipyretics and inhaled medicines are in main shortage, but Greek pharmacists noted that more than 200 drugs for almost the entire spectrum of diseases were in permanent shortage.

The Causes
It is noted that 60-80% of the products used in medicines are produced in China and India. Their production has fallen by 60% due to a new outbreak of the coronavirus in China resulting in limited exports. Another reason is the war in Ukraine delaying distributions main reason for drug shortages. Unfortunately, Europe has chosen to depend to a significant extent on China and India in the field of pharmaceutical raw materials. According to high-ranking officials in the sector, the objective of the European Commission is to “move” production from Asian countries to Europe, in order to facilitate the supply of medicines within the continent. Of course, in this particular period of time, with inflation soaring, and the energy crisis exponentially increasing the cost of drug production, a feeling of insecurity is being cultivated as to whether Europe will be able to respond to a new wave of shortages of even more important drugs.

Another cause according to pharmacists, is the ever-increasing parallel exports. That’s because certain medicines can give up to 15 times more profit to the pharmaceutical warehouse if it is available abroad than in a pharmacy in the Greek market. The National Pharmaceutical Organization of Greece (EOF) decided to put a brake on the parallel exports of a large category of medicines for which there are shortages in the Greek market. In addition to the previous measures, the Greek Minister of Health asked the warehouses to immediately declare the stocks they have of the elliptical drugs and they must make them available without delay to the Greek market.

Nevertheless, reports claim that of the drugs whose export was banned on November 22, 80% of them have identical generics, so there is no public health issue.

?he solution that consumers refuse due to unawareness
Generic medications are a class of medication that, while lacking the brand name, are identical to brand-name products in terms of dosage, strength, administration method, quality, and intended use. They are frequently accessible after the original drug’s patent has expired, enabling other pharmaceutical companies to create and market less expensive versions of the drug. The primary motivation behind the development of generic medications is to lower the cost of healthcare. The pharmaceutical business that created the new medicine received a patent for it. This patent grants the corporation the sole right to market the medication for a specified period of time. Because there are no other options during this time, the manufacturer can charge a premium price for the drug. When the patent on the drug expires, other businesses may begin making and marketing generic versions of it. Because the firms making these generic versions do not have to invest in the research and development that went into manufacturing the original drug, they are often far less expensive than their brand-name counterparts. Drugs sold under generic names are as secure and efficient. They function similarly to the original medication and have the same active components. Generic medications must go through extensive testing to guarantee that they are identical to the brand-name medication in terms of quality, safety, and effectiveness. While using generic medications can help reduce the cost of healthcare, there are some instances where the brand-name choice may be chosen.

For instance, if a patient has previously experienced a negative reaction to a generic version of a medication, their doctor may advise that they take the brand-name version to prevent any prospective problems. Finally, we understand that if consumers are made aware of what generics are and of the controls that must be submitted to be released on the market, in periods of shortages we will not face such a serious issue in treatments.

Georgios-Marios Bolmpasis
MPharm Student University of Athens
Vice President of Pharmacy Students Association(Athens)
Member of European Pharmaceutical Students Association
Member of Greek Pharmaceutical Students Federation

Read More

Source Here: pharmatutor.org

Continue Reading

Pharma

Computational Analysis of HCV Entry Inhibitors for Hepatitis C Treatment – a Molecular Docking Approach

Yesenia Harris

Published

on

By

About Author: Karthika M
Postgraduate in Bioinformatics
Bharathiar Universitym.karthika1@gmail.com

ABSTRACT
Hepatitis C virus (HCV) is a major human pathogen associated with life-threatening liver disease, affect 170 million people worldwide. Hepatitis C virus (HCV) envelope proteins mediate the entry of virus into cells by binding to cellular receptors, resulting in fusion of the viral membrane with the host cell membrane and permitting the viral genome to enter the cytoplasm. Entry inhibitors would inhibit the viral entry itself.
Envelope protein E2, responsible for viral entry taken as target is modeled using Modeller 9.10. 47 small molecules have been taken for docking with envelope protein.10 small molecules have been selected for Induced Fit Docking based on the Glide energy, Docking Score.
Ligand 20 is showing good binding affinity towards the target in active site, having Docking Score -6.07, Glide Energy -60.41, IC50 value 1.5 and it obeys Lipinski’s Rule of Five. ADMET Properties were also studied for these compounds. All computational works are performed in the Schrodinger suite 2009. Hence the present study suggests that the ligand 20 with further in vitro and in vivo testing can be introduced as an effective inhibitor against HCV for development of curative therapy against Hepatitis C.

REFERENCE ID: PHARMATUTOR-ART-1650

INTRODUCTIONHepatitis C is a contagious liver disease that results from infection with hepatitis C virus (HCV). The infection is often asymptomatic, but chronic infection can lead to scarring of the liver and ultimately to liver cirrhosis. In some cases, those with cirrhosis will go on to develop liver failure, liver cancer or life-threatening esophageal and gastric varices. HCV is transmitted through contact with the blood of an infected person. (Jou, Muir, 2008)

The Hepatitis C virus persists in the liver in about 85% of those infected. This persistent infection can be treated with medication; peginterferon and ribavirin are the current standard therapy. Overall, between 50-80% of people treated are cured. Those who develop cirrhosis or liver cancer may require a liver transplant. Hepatitis C is the leading cause of liver transplantation though the virus usually recurs after transplantation. No vaccine against hepatitis C is currently available.(Ghany MG et al.,2009)

Hepatitis C virus(HCV)
Hepatitis C virus (HCV) is a small (55-65 nm in size), enveloped, positive-sense single-stranded RNA virus of the family Flaviviridae. Hepatitis C virus is the cause of hepatitis C in humans.(Kapoor et al.,2011)

Structure

Fig : Structure of Hepatitis C Virus

The hepatitis C virus particle consists of a core of genetic material (RNA), surrounded by an icosahedral protective shell of protein, and further encased in a lipid (fatty) envelope of cellular origin. Two viral envelope glycoproteins, E1 and E2, are embedded in the lipid envelope.

Genome

Fig: HCV Genome

HCV Envelope Protein as a Target
Cellular processes, such as clathrin-mediated endocytosis or endosome formation, are probably not very clean targets, and agents designed to target those processes would be likely to have severe adverse effects. Agents that bind host factors may interfere with normal cellular functions, resulting in an unacceptable adverse effect profile. Agents that bind viral components in the entry process, such as the HCV envelope proteins, might be more likely to be selected as target.

Entry Inhibitors
Entry inhibitors may be particularly helpful after liver transplantation, where universal reinfection of the graft often results in rapid fibrosis progression and subsequent graft failure. Even therapies transiently inhibiting HCV cell entry could prevent graft reinfection and thus greatly improve the effectiveness of liver transplantation.

MATERIALS AND METHODS

Molecular Modelling
This procedure, also termed Comparative Modelling or knowledge based modelling developed a three-dimensional model from a protein sequence (target) based on the structures of homologous proteins (Templates).

Modeller
Modeller is a computer program that model three-dimensional structures of proteins and their assemblies by satisfaction of spatial restraints.

The user provided an alignment of a sequence to be modeled with known related structures and Modeller automatically calculated a model containing all non-hydrogen atoms.More generally, the input to the program was restraints on the spatial structure of the amino acid sequence and ligands to be modeled. The output was a 3D structure that satisfied these restraints as well as possible. A 3D model was obtained by optimization of a molecular Probability Density Function (PDF). The molecular PDF for comparative modeling was optimized with the variable target function procedure in Cartesian space that employed methods of conjugate gradients and molecular dynamics with simulated annealing.

Searching for Structures
First, it was necessary to put the target sequence into the PIR format readable by Modeller.

A search for potentially related sequences of known structure could be performed by the profile.build() command of MODELLER.

Selecting a Template
To select the most appropriate template for the query sequence over the similar structures, we would use thealignment.compare_structures() command to assess the structural and sequential similarity between the possible templates.

Model Building
Once a target-template alignment was constructed, MODELLER calculated a 3D model of the target completely automatically, using its automodel class. The following script would generate five similar models of HCV E2 based on the template structure and the alignment file “align_HCV.ali”. The most important output files were ‘model-single-log’, which reported warnings, errors and other useful information including the input restraints used for modeling that remain violated in the final model, which contained the coordinates of the five produced models, in the PDB format.

Model Evaluation
If several models were calculated for the same target, the ‘best’ model could be selected by picking the model with the lowest value of the MODELLER objective function. The DOPE potential in MODELLER was used to evaluate the model fold.

Molecular Docking
The computational process of searching for a ligand that is able to fit both geometrically and energetically to the binding site of a protein is called molecular docking.

Docking Analysis using MAESTRO
Maestro is the graphical user interface for all of Schrodinger’s products like CombiGlideTM, EpikTM, GlideTM, ImpactTM, LiaisonTM, LigprepTM,MacroModelTM,PhaseTM, PrimeTM, QikPropTM, QsiteTM, and StrikeTM. It contains tools for building, displaying, and manipulating chemical structures for organizing, loading and storing these structures and associated data, and for setting up, monitoring, and visualizing the results of calculations on these structures.

Docking Method – GLIDE(Grid Based Ligand Docking with Energitics)
Glide searches for favorable interactions between one or more typically small ligand molecules and a typically larger receptor molecule usually a protein. Each ligand must be a single molecule, while the receptor may include more than one molecule such as a protein and a cofactor. GLIDE can be run in rigid or flexible docking modes and it the later automatically generates conformation for each input ligand. The combination of positions and orientation of the ligand relative to the receptor along with its conformation in flexible docking is referred to as a ligand pose.

Protein Preparation
A typical PDB structure file consists only of heavy atoms. Therefore, hydrogen does have to be added prior to use in Glide calculations, which use an all-atom force field.

The protein preparation facility consists of two components, preparation and refinement. After ensuring chemical correctness, the preparation component adds hydrogen and neutralizes side chains that are not close to the binding cavity and do not participate in salt bridges. The refinement component performs a restrained impact minimization of the co-crystallized complex, which reorients side-chain hydroxyl groups and alleviates potential steric clashes.

Ligand Preparation
Glide also allows importing the lead molecule in SDF, mol or mol2 format which is drawn using chemsketch software. These determined structures are minimized using two methods
1. Ligprep Minimization
2. Impact Minimization

Ligprep generate tautomers and conformers for single ligand and Impact minimization uses two algorithms for minimization steepest descent and conjugate gradient which runs for 500 to 1000 cycles. Both methods use OPLS force field for minimizing the structures.

Receptor Grid Generation
The ligand should be selected for the ligand to identify the active site to bind with the protein.

If the protein structure in PDB is not complexed with ligand, then the active site of the protein should be specified. We can particularly specify the residues or can specify the region.

Ligand Docking
For docking the ligand with the target protein the receptor grid file should be specified and the ligprep out file should be selected.

Designing of Inhibitors
Novel class of HCV Rimcazole derivatives were recently reported for their anti-HCV activity and they were designed using MARVINSKETCH and PYMOL. Totally 47 Rimcazole derivatives were modeled using MarvinSketch software.

Collection of protein
Protein is retrieved from PDB (protein data bank) based on the lowest resolution. To model the protein, the template structure is selected (PDB id : 2xfc) from PDB.

Modelled Structure of HCV E2

Protein Information

Length

862 Residues

Target Sequence

Uniprot ID – Q99IB8

Template

Chikungunya Virus Envelope Protein E2 PDB ID – 2XFC

Active Site Residues

Lys 373, Asn 409, Trp 414, Asp 417, Glu 418, Leu 705, Tyr 706, Asp707, Ser 708, Ser 710, Thr 712, Val 714.

RESULTS

Ramachandran Plot for HCV E2 Glycoprotein

Docking Result

Compounds

Docking Score

Glide Energy(kcal/mole)

20

-4.01

-51.50

19

-5.32

-49.91

34

-5.35

-49.75

6

-5.51

-49.72

7

-4.8

-49.65

4

-6.24

-48.84

3

-6.24

-48.84

5

-6.24

-48.84

30

-4.49

-48.28

27

-3.92

-47.32

8

-5.08

-46.68

22

-4.74

-46.56

12

-4.01

-46.55

28

-4.40

-46.28

33

-5.28

-46.20

35

-4.82

-45.55

18

-4.58

-45.39

14

-4.69

-45.21

41

-4.83

-44.95

9

-4.88

-44.63

26

-3.62

-44.62

46

-4.55

-44.51

44

-4.55

-44.51

45

-4.55

-44.51

43

-4.55

-44.51

47

-4.55

-44.51

42

-4.55

-44.51

31

-5.32

-44.21

23

-5.26

-43.89

32

-4.44

-42.59

10

-4.12

-41.70

40

-6.58

-41.40

Induced Fit Docking Results

Ligands

Docking Score

Glide Energy (kcal/mole)

Interaction

Distance (?)

20

-6.07

-60.41

[N-H…O] Tyr367

Lys373[N-H…O]

3.043

2.986

25

-6.84

-56.73

Lys373[N-H…O]

Lys574[N-H…O]

3.017

2.803

30

-8.19

-55.52

Asn409 [N-H…O]

Gln418 [N-H…O]

3.022

2.848

24

-6.08

-54.17

Lys373[N-H…O]

Lys574[N-H…O]

2.918

3.089

34

-7.85

-54.01

[N-H…O] Asp417

Asp417 [N-H…O]

Asn409 [N-H…O]

Trp414 [N-H…O]

3.796

3.130

2.867

2.852

Induced Fit Docking Results of Ligand 20

Poses

Docking Score

Glide Energy (kcal/mole)

Interaction

Distance (?)

1

-6.07

-60.41

[N-H…O] Tyr367

Lys373[N-H…O]

3.043

2.986

2

-5.70

-55.90

Lys373 [N-H…O]

Lys371 [O-H…O]

2.980

3.044

3

-4.26

-54.70

Lys373 [N-H…O]

Lys574 [N-H…O]

[N-H…O] Glu577

2.992

2.802

4

-8.19

-55.52

Asn409 [N-H…O]

Gln418 [N-H…O]

3.022

2.848

5

-6.30

-54.42

[N-H…O] Glu577

Lys574 [N-H…O]

Lys373 [N-H…O]

2.909

3.065

2.732

6

-7.44

-53.27

Gln418 [N-H…O]

2.895

7

-7.62

-53.13

Gln418 [N-H…O]

Asn409 [N-H…O]

2.904

3.214

Interaction of Ligand 20 in PYMOL

Ligplot of Ligand 20

DISCUSSION
The compound 20 is most potent with an IC50 of 1.5 nM because of the presence of methyl ester group in the series of compounds. The methyl ester at the 3-position improved the potency by several fold. The dimethyl sulfonamide compound 15 showed excellent potency with an IC50 of 5 nM. The N-methylated analogue 24 is as potent as compound 20. When compared to the corresponding 4,6-disubstituted-2-amino pyrimidines, aromatic heterocycles/aromatics such as phenyl sulfone (30) or difluoro phenyl sulfonamides (34) are also active but less potent than pyrimidine analogues. Disubstituted 2-amino/2-oxo pyrimidines and dimethyl sulfonamide groups are best for high potency. Based on binding competition and resistance profiles, this compound class appears to be mechanistically similar to ITX 5061.

CONCLUSION
Envelope Protein E2 (Glycoprotein) responsible for HCV Entry is targeted to inhibit the HCV. Both protein and ligands are minimized using OPLS forcefield. From the 47 compounds, 10 compounds were selected for Induced Fit Docking based on Docking Score and Glide Energy. The ligands chosen abides Lipinski’s Rule of Five. These small molecules belong to a distinct chemical class from ITX 5061, first in class drug, currently in Phase Ib Clinical Trial.

Ligand 20 is showing good binding affinity towards the target having Docking Score -6.07 and Glide Energy -60.41 and IC50 value 1.5nM and it binds with the Active site of the target protein. Hence the present study suggests that the ligand 20 with further in vitro and in vivo testing can be introduced as an effective inhibitor against HCV for development of curative therapy against Hepatitis C.

References
1) Kato N (2000). “Genome of human hepatitis C virus (HCV): gene organization, sequence diversity, and variation”. Microb. Comp. Genomics 5 (3): 129-51.
2) Op De Beeck A, Dubuisson J (2003). “Topology of hepatitis C virus envelope glycoproteins”. Rev. Med. Virol. 13 (4): 233-41.
3) F. Habersetzer, A. Fournillier, J. Dubuisson, D. Rosa, S. Abrignani, C. Wychowski, I. Nakano, C. Tr?po, C. Desgranges, G. Inchausp?, Characterization of human monoclonal antibodies speci?c to the hepatitis C virus glycoprotein E2 with in vitro binding neutralization properties, Virology 249 (1998) 32-41.
4) Targeting HCV entry for development of therapeutics. Wong-Staal F, Syder AJ, McKelvy JF.Viruses. 2010 Aug;2(8):1718-33. Epub 2010 Aug 18.
5) Glycosylation of hepatitis C virus envelope proteins. Goffard A, Dubuisson J. CNRS-UPR2511, Institut de Biologie de Lille, Institut Pasteur deLille, Lille, France. Biochimie. 2003Mar-Apr;85(3-4):295-301.
6) Z.Y. Keck, A. Op de Beeck, K.G. Hadlock, J.M. Xia, T.K. Li, J. Dubuisson, S.K.H. Foung, Hepatitis C virus E2 has three immunogenic domains containing conformational epitopes with distinct properties and biological functions, J. Virol. 78 (2004)9224-9232.
7) G. Leroux-Roels, R. DeLeys, L. Stuyver, A.Elewaut, J. Philipp?, I. Desombere, J.Paradijs, G. Maertens, Lymphoproliferative responses to hepatitis C virus core, E1, E2, and NS3 in patients with chronic hepatitis C infection treated with interferon alfa, Hepatology 23 (1996) 8-16.
8) B. Bartosch, J. Dubuisson, F.L. Cosset, Infectious hepatitis C virus pseudo- particles containing functional E1-E2 envelope protein complexes, J. Exper.Med. 197 (2003) 633-642.
9) M. Rodriguez-Rodriguez, D. Tello, B. Yelamos, J. Gomez-Gutierrez, B. Pacheco, S. Ortega, A.G. Serrano, D.L. Peterson, F. Gavilanes, Structural properties of the ectodomain of hepatitis C virus E2 envelope protein, Virus Res. 139 (2009) 91-97.
10) V. Sandrin, P. Boulanger, F. Penin, C. Granier, F.L. Cosset, B. Bartosch, Glycosylation of hepatitis C virus envelope proteins, Biochimie 85 (2003) 295-301.
11) A.OpDe Beeck, L. Cocquerel, J. Dubuisson, Biogenesis of hepatitis C virus glycoproteins, J. Gen. Virol. 86 (2005) 3189-3199.
12) A. Op De Beeck, C. Voisset, B. Bartosch, Y. Ciczora, L. Cocquerel, Z. Keck, S. F. Helle, J. Dubuisson, Hepatitis C virus entry into host cells,Cell Mol. Life Sci. Cell entry of hepatitis C virus, Virology 348 (2006) 1-9.
13) M. Lambot, S. Fretier, A.O. De Beeck, B. Quatannens, S. Lestavel, W. Clavey, J. Bartosch, B.; Dubuisson, J.; Cosset, F.L. Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes. J. Exp. Med. 2003, 197, 633-642.
14) Bartosch, B.; Dubuisson, J.; Cosset, F.L. Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes. J. Exp. Med. 2003, 197, 633-642.
15) Bankwitz D., Steinmann E., Bitzegeio J., Ciesek S., Friesland M., Herrmann E., Zeisel M. B., Baumert T. F., Keck Z. Y., et al., Hepatitis C virus hypervariable region 1 modulates receptor interactions, conceals the CD81 binding site, and protects conserved neutralizing epitopes. J Virol 84, 5751-5763.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

Read More

Source Here: pharmatutor.org

Continue Reading

Trending

RedFoxNews.com