Process For Preparation Of Spray Dried Feracrylum And Composition Comprising The Same

Abstract

A process for Feracrylum powder by spray drying such that the feracrylum powder produced is of fine dispersibility with acceptable quality parameters and a particle size distribution of below 100μ, said process comprising providing feracrylum solution: supplying air through micron filters and heating at 135ºC to 145ºC; spraying Feracrylum in a drying chamber kept at 140ºC and vacuum is maintained at-20 to -30 mm WC; collecting Feracrylum in bag filter at a filter pressure of 5-10 mm; maintaining nozzle air pressure for atomization at 1.5 to 2.5kg/ Cm2 and feed rate of the solution at 50 m1/min to 100 ml/min. the invention also relates to a comprising 1 to 12% w/v Spray dried Feracrylum having particle size below 100, prepared by the process comprising providing feracrylum solution; supplying air through micron filters and heating at 135ºC to 145ºC; spraying Feracrylum in a drying chamber kept at 140ºC and vacuum is maintained at -20 to-30 mm WC; collecting Feracrylum in bag filter at a filter pressure of 5-10 mm; maintaining nozzle air pressure for atomization at 1.5 to 2.5 kg/Cm2and feed rate of the solution at 50 ml/min to 10 ml/min: and pharmaceutically acceptable excipients, the composition having a pH of 2.5-6.5

Information

Applicants

Inventors

Specification

FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)
1. Title of the invention: PROCESS FOR PREPARATION OF SPRAY DRIED FERACRYLUM AND COMPOSITION COMPRISING THE SAME
2. Applicant(s)
(a) NAME : PATEL, Dinesh, Shantilal ;
PATEL, Sachin, Dinesh ;
KURANI, Shashikant, Prabhudas
(b) NATIONALITY : Indian
(c) ADDRESS : THEMIS MEDICARE LIMITED, 11/12,
Udyognagar, S V Road,'Goregaon (W) Mumbai -400 104, State of Maharashtra, India.
3. PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in which it is to be performed:

Field of Invention
The present invention is concerned with process for preparation of Pharmaceutical grade Feracrylum for preparation of various pharmaceutical compositions and
galenicals. More specifically the present invention relates to the process for preparation based on Spray dried powder of Feracrylum with quality specifications for better dispersion of locally delivered Feracrylum through formulations in the form of solutions, powders, lotions, spray, paste and gels, creams for local, oral, ear, nasal or bronchial application. The present invention further relates to composition comprising Feracrylum prepared by the method of present invention.
The present invention also provides a method of treatment of wounds which may or may not be bleeding as well as method of treatment of disease or conditions which are associated with blood loss like haemorrhoids in animals and humans as well for other reasons.
BACKGROUND AND PRIOR ART :
Numerous organic compounds are poorly soluble or water insoluble and increasing the surface area, increases the solubility in the media. It is therefore, necessary to find procedures for dissolving such compounds sufficiently well in aqueous system because otherwise their efficacy is greatly impaired. Poorly soluble Active Product ingredient sometimes make it difficult to come in contact with surfaces where such matters are coming directly in contact with the mucosal surface area thereby reducing the efficacy. Various possibilities are already known for improving the solubilization and dispersion in aqueous or other media by reducing the particle size for poorly soluble or dispersible substances.
US 4,215,106 teaches the use of Feracrylum as a haemostatic. This patent describes the product Feracrylum and, the methodology and description employed for synthesis of this product, which can be termed as an industrial product and not a pharmaceutical product, because no standard or specification of the product has been reported. A pharmaceutical preparation requires specification and method of testing. Moreover the
product of US Patent 4,215,106 is always obtained in a plastic-like material. The
2
Feracrylum prepared by this patented process is obtained as waxy flakes in nature and
is difficult to powder.
Further PCT Application No. WO 01/74370 (confirmed) describes novel
pharmaceutical preparation of Feracrylum for the treatment of peptic and duodenal ulcers caused by H.pylori and cholera which is caused by Vibrio cholera claiming it as antibacterial/antiprotozoal in the form of tablets / syrups and other prepared formulations.
This art also describes preparation of Feracrylum in different ratios then what is described in the US Patent 4,215,106 (please clarify what is referred by different ratios). However, it fails to reach the stringent quality specification which would be more acceptable to the pharmacist for their pharmaceutical compositions. In US 4,215,106 patent, difficulties encountered by formulators were resolved to large extent in the later PCT patent however it also failed to deliver Pharma grade Feracrylum. Even after this improvement there is consistent need for the improvement of quality.
In US Patent 4,215,106 the material obtained was Brownish in colour and in WO 0174370 material obtained was a Peach coloured. No statement with regards to quality or particles are ever stated in US 4,215,106. The particles were controlled upto 700 u in WO 01/7437.
In order to achieve properties, eg. Contact of mucosal surfaces, it is necessary for poorly soluble or difficultly soluble compounds to be dispersed as finely as possible
in matrices. A particle size of less than 100p. is desirable for compounds like Feracrylum. Such particle was difficult to be achieved by crushing or by milling of polymers. The grinding may lead to uneven distribution. Attempts have been made in past for doing such in WO 01/74370 by keeping the particles in the size range of 700 p. particle size but still dispersibility could not be achieved, though specifications are meeting Q.C. requirements. The result of dispersed matter in matrices showed poor Bioavailability to the mucosal surfaces. No such attempt for evaluating was ever made by prior US 4,215,106 for this compound. While trying to achieve the material using
process of US 4,215,106 patent process has resulted in a Feracrylum as lumps not meeting quality standard. Therefore, there is consistent need to achieve the material
3

which not only meets Q.C. but also useful in terms of easy dispersibility of medicaments for better efficacy.
The control of particle size is very important for application in formulation with respect to easy dispersibility and dissolution of the material. These characteristics are absent in the US 4,215,106. The WO 01/74370 though controlled the colour of Feracrylum but has failed to control the particle size.
Further the process of prior art involves evaporation of liquid Feracrylum thereby forming coloured lumps and difficult to remove Feracrylum from surfaces where the process of drying is carried out. Thus some times requiring physical and manual efforts by scratching of the surface of vessel or tray/paper. If, it was to be dried on parchment paper it results in contamination of fibres and other unknown material. This also provides an impure product mixed with contaminant requiring further filtration before application for surgical wounds. Hence not recommended.
Further during the synthesis of polymer removal of liquid during the synthesis as cited in US 4,215,106/ WO 01/74370 leads to lumps or flakes respectively and has posed difficulties and disadvantages on one hand because it represents an energy spending process which must take place at elevated temperature and with reduced pressure thereby causing increase in colour and formation of lumps because of thermal stress and even slight contamination during the process.
Further in the above prior art the compounds are very hygroscopic and the material with higher hygroscopic quality leads to lumps and shows physical instability during the storage and formulation.
All the above problem required improvements in terms of physical characteristic like quality, particle size, colour and dispersibility.
The problems which have been mentioned make it clear that despite the advantages of the process as described primarily, economic and efficacious production of
Feracrylum was always difficult as it is not complying with quality standards as framed in GMP requirements and for pharmacists acceptability of formulations.
4

Accordingly there is need to provide Feracrylum which satisfy the following conditions: l)appropriate description 2) to produce desired effect on adding as to
dispersion 3)to be effective after its dispersion and easy utility 4) and easily applicable
formulations.
Objects of the Invention
Thus one object of the present invention is to provide a process of producing a Feracrylum of a acceptable quality containing microparticle of Feracrylum which can be easily dispersed in a various galenicals like tulle, Spray, Gauzes, Creams, tablets, capsules to the advantage over prior arts.
Another object is to prepare Feracrylum free from all above cited problems and prepare Feracrylum which can control colour, hygroscopy and contamination leading to good stability, dispersibility and efficacious products.
Further object of the present invention is to provide micronised spray dried powder of Feracrylum with stringent quality specification of particular size of below l00μmsuch that the formulations prepared by such quality material has lead to better formulation in terms of dispersibility, easy solubility, spreadibility and efficacy.
Summary of the Invention
The invention therefore relates to a process for phramaceutically acceptable Feracrylum powder by spray drying such that the feracrylum powder produced is of fine dispersibility with acceptable quality parameters and a particle size distribution of below l00μ, said process comprising
i) providing feracrylum solution;
i) supplying air through micron filters and heating at 135°C to 145°C;
ii) spraying Feracrylum in a drying chamber kept at 140°C and vacuum is
maintained at -20 to -30 mm WC;
iii) collecting Feracrylum in bag filter at a filter pressure of 5 - 10 mm with outlet
temperature of chamber kept between 80°C to 90°C preferably at 85°C
5
v) maintaining nozzle air pressure for atomization at 1.5 to 2.5 kg/cm2and feed rate of the solution at 50 ml/min to 100 ml/min
According to another aspect there is provided a composition comprising 1 to 12% w/v Spray dried Feracrylum having particle size below 100μ, prepared by the process comprising
i) providing feracrylum solution;
ii) supplying air through micron filters and heating at 135°C to 145°C;
iii) spraying Feracrylum in a drying chamber kept at 140°C and vacuum is
maintained at -20 to -30 mm WC;
iv) collecting Feracrylum in bag filter at a filter pressure of 5 - 10 mm with outlet
temperature of chamber kept between 80°C to 90°C preferably at 85°C
v) maintaining nozzle air pressure for atomization at 1.5 to 2.5 kg/cm2and feed rate of the solution at 50 ml/min to 100 ml/min and pharmaceutically acceptable excipients, the composition having a pH of 2.5-6.5
Detailed description of Invention
The process of present invention provides feracrylum with easy dispersibility and hence prolonged shaking efforts are avoided during the stages of formulation.
The invention therefore relates to process for producing Feracrylum of fine dispersibility with acceptable quality parameters which comprises of production of Spray dried powder by a process which is economically beneficial. The Feracrylum thus produced has Quality specification for achieving the pharmaceutically acceptable Feracrylum for compounding and is efficacious in terms of easy availability in contact with the tissues for haemostatic/antibacterial activity.
The process additionally provide economic production of pharmaceutically acceptable quality of Feracrylum as given in the specification without impurities.
This invention also provides a composition of Spray dried powder of Feracrylum formed by the process of present invention-which is normally not antigenic and is not absorbed in the systematic circulation presumably because of the size of the molecule.
4

The process of Spray Drying is carried out as follows using the economically viable instrument.
The ambient air enters through the air filter and is sucked into the system by the supply
fan. The air is heated in steam radiator and sent into the Spray drier through the air disperser.
Closed feeder tank is loaded with Feracrylum as received from the synthetic process at the flow of 50 ml/min to 100 ml/min at 10% to 20% of feed stroke into feeder tank. The feed from the feed tank is fed to the Spray nozzle by a reciprocating plunger pump. Compressed air is used for atomization of the Spray into fine droplets.
The atomized Spray and the hot air mix in the drying chamber. Primary separation of the powder takes place in the cyclone. The exhaust air is sent to a bag filter, where secondary separation of fines is effected.
The dried powder is collected under the cyclone and the bag filter, through a flap valve to powder collector pot underneath. The clean exhaust air is then sent to the stack
though exhaust fan.
The Spray dried Feracrylum so obtained has a controlled quality specification with a particle size distribution of not more than l00μ, preferably 60-70 μm.
The Spray dried material has easy dispersion in water while dissolving in water leads to easy dispersibility with a colour index of 0.01 - 0.2 when determined at 435 nm.
Further such a Feracrylum as prepared has better dispersibility in a matrices of gel, tulle and other formulation exhibiting better efficacy in terms of healing as against Feracrylum prepared as described in WO 01/74370.
Further this process maintains the product sterility without contamination in terms of microbial load. It is very important for a product which is used for surgical application for treating the wound in a conditions whether as a haemostatic or antiseptic. This process of Spray drying ensures the above requirement of product sterility which is not achievable by US 4,215,106.
7

This type of Spray drying gives an anhydrous material with a quality specification which meets the laid down specification and good control over particle size. This
material when dissolved in water has a colour index from 0.01 to 0.2 unlike the colour index of Feracrylum produced by US 4,215,106 and WO 01/74370. In the product formed by the present process alongwith the dispersibility the efficacy in terms of wound healing is higher than the product obtained by US 4,215,106 and WO 01/74370.
The Spray drying plant is designed specifically to ensure that there is no contamination of material from the external source. All the contact parts of air, feed and product are in stainless steel 316.
Incoming air is filtered through pre-filtered followed by HEPA filter. For atomization also a two fluid nozzle using compressed air is used. The compressed air is free from oil free compressor which ensures of no oil contamination of the product. In contrast, rotary atomizer uses oil for lubrication of bearings which can contaminate the product. All the flanges for the ducts are grooved, silicone rubber gaskets are used, thus the plant can be sterilized at high temperature, prior to product drying. A tight seal butterfly valve is used as an air lock valve, instead of a rotary valve, which could be a source of contamination.
The process itself is very easy to operate with an efficiency in the product collection. There are minimum moving parts in the plant. The two fluid nozzle used for atomization is easy to assemble and clean, with no moving parts as compared to a high speed rotary atomizer, which operates at an rpm of 20,000. The feed pump is a plunger pump running at low rpm, ensuring minimum wear and tear. The exhaust blower is designed to operate at a low rpm.
This Spray drying system makes it easier to clean the whole system without the residual contamination.
The powder is primarily collected in a high efficiency cyclone collector. For collection of the fines leaving the cyclone separator, a pulse jet bag filter is used. The bags are
8

with Teflon membrane lining, ensuring surface filtration and negligible losses to the atmosphere. The chamber is provided with a cleaning door of adequate size for through cleaning. The ducts are provided with grooved type flanges at regular intervals, for cleaning. The pulse jet bag filter cover is provided with a davit arrangement for easy access and removal of the bags.
Further it gives a aesthetic look. The entire plant including the supporting structure is in stainless steel. This gives a pleasing look to the plant.
The quality specifications of such Spray dried powder obtained by process of present invention is as follows:

Description Cream coloured powder
Solubility 1 - 12% in water
pH [1% aq. Solution] 2.4 to 4.0
Relative Viscosity [at 25 °C] 2 cps to 5 cps
Loss on drying Max. 5%
Heavy metals N.M.T. 0.002%
Iron 0.1% to 0.4%
Unreacted protein Max. 10%
Colour & clarity of solution 0.01 to 0.2
Activity of Feracrylum Min.90%
Bulk Density 0.2 to 0.5 g/cc
Particle Size Less than 100μ
The product so obtained has an advantage with a solubility and dispersibility profile better than WO 01/74370 and US 4,215,106.
Further the physical properties of the spray dried powder of present invention differs from that of the prior art methods. The said properties are tabulated below:
9

Sr. No. Tests US Patent WO 0174370 Present Invention
1) Description Brownish sticky Granular peach Creamish fine
flakes coloured powder powder
2) Colour Index at 435run for 1% soln. In 1 cm cell A = 0.362 A = 0.2 A = 0.08
3) Particle Size More than 1000 Average particle Average 60 -
um, difficult to size is 500 urn 70 um
sieve as is and waxy pasty
4) Loss on Drying 12% 4.3% 2.5%
Composition according to the present invention comprises dispersible macromolecular spray dried powder of Feracrylum formed by the process of present invention comprising particles having average particle size below l00μ preferably 90% of Feracrylum comprises of size in the range of l00μ
Various formulations made from Spray dried Feracrylum prepared which includes sterile liquid form and preparations made thereof in aqueous media with propellant and solid forms.
The liquid form may be a solution, suspension or other dispersion of Feracrylum in a suitable liquid carrier, preferably in a liquid solvent in combination with pharmaceutically acceptable excipients. The ideal liquid carrier may be water. It is possible to incorporate organic solvents like ethanol and alike in the spray form.
The formulation of Feracrylum for oral application comprises of dispersion of Feracrylum with sugar, colourant, the tablet formulation prepared from Spray dried Feracrylum in composition with gelatin, starch, PVC, PVP, methyl cellulose, CMC, HPMC and for Feracrylum gels which also can contain alginate and coolant like menthol, stabilizers like parabens and polymers expanding material for better spreadability, anaesthetics like lignocaine, centbucridine, bupivaccaine, tetracaine, antibiotics like polymixin derivaties, soframycin, neomycin, clindamycin, analgesics of NSAID eg. diclofenac, aceclofenac, piroxicam, tramadol, ibuprofen, indomethacin, naproxen, antiseptics like povidone iodine, triclosan, antibacterial like nitroimidazole, metronidazole, secnidazole or its derivatives and antifungals like azoles or allylamines
10

eg. clotrimazole, fluconazole, itraconazole, terbinafine, natural products like curcumin or products of Aloe barbadensis and other natural products having antibacterial, antifungal activity, antipruritic substances like diphenhydramine HC1, Calamine,
various steroids alone or in combinations.
The concentration of Feracrylum may vary from 1% w/v to 12% w/v of an aqueous composition which composition may optionally contain other adjuvants.
Spray dried Feracrylum may be prepared in solution form, in various concentrations, preferably from 1% - 12% w/v or w/w depending on the formulation. The solution prepared is limpid, stable, safe and effective for topical application. The amount of water can be up to 89% v/v by volume of said composition.
Further Spray dried Feracrylum so prepared as above is dispersed in PEGS to give a fine dispersion which can be spreaded on cotton gauze or rayon gauze and cut into pieces and placed between a parchment paper and placed in a suitable container to give ready to use gauze after y ray sterilization.
The liquid dispersion of the compounds is made in distilled water in the range of 1% to 5% by weight and such formulation may be applied directly to wounds or bleeding surfaces for stopping the bleeding by applying a liquid so prepared using a sterile ear buds for clotting of blood and providing a surface protection for bacterial attack. Feracrylum also helps in wound healing because of its inherent antibacterial property.
The composition comprising of Spray dried Feracrylum is advantageously used as an antimicrobial and antiseptic agent and optionally as a haemostatic agent.
The composition may be in the form of solution, spray, powder, ointment, cream, gel, lotion, suppository and the like. The pH of the liquid composition is maintained between 2.5 and 6.5 and preferably between 2.9 to 4.0.
For oral dosages, the Feracrylum composition may include tableting adjuvants, including pharmaceutically acceptable tableting adjuvants selected from fillers, excipients, compression aids, binders, flavorings, coating agents, and the like.
II

The composition comprising Spray dried Feracrylum can be formulated in a gel with conventional components such as gum tragacanth, sodium alginate, methyl cellulose,
sodium carboxy methyl cellulose, carbomers, polyvinyl alcohol, propylene glycol,
cetostearyl alcohol, glycerol, benzyl alcohol, isopropanol, titanium dioxide, butylated hydroxy toluene, disodium edetate, etc; as a lotion with conventional components such as polyethylene glycols-400, butylated hydroxyanisole, white soft paraffin, cetostearyl alcohol, propylene glycol, methyl paraben, propyl paraben, as a powder with conventional components such as medicated talc, titanium dioxide, etc.; or as a cream with conventional components such as white soft paraffin, liquid paraffin, cetomacrogol - 1000, cetostearyl alcohol, water, chlorocesol, carbomers propylene glycol, titanium dioxide, etc.
A Feracrylum solution can be filled under sterile conditions in foam filled packing, in volumes varying from 5 ml to 500 ml. The formulation may be prepared in single and multi-dose bottles and also in the form of 'Spray' and 'pre-filled' syringe for direct use in clinical practice.
Haemorrhoids are very common in animals and humans and when a suppository containing Spray dried Feracrylum can be used in form of gels containing anaesthetics like Centbucridine Hydrochloride, Lignocaine it is found to prevent bleeding by relieving the painful condition.
Spray dried Feracrylum may be used directly or in the form of aqueous solution, which can be spread on cotton gauze, and the latter is applied on to a bleeding surface to arrest bleeding. The wounds may arise out of accidents, surgery and other reasons. The composition of the invention is applied to the site of wound. It is generally observed that surgeons, while operating, are constrained for time and procedures of this application viz, use of gauze are very lengthy. The procedure involves spreading of the Feracrylum solution on gauze or cotton wool tampons, applying the same to a bleeding surface and then waiting for some time until bleed is arrested.
The composition of the invention of solution of Feracrylum for irrigation during the surgery overcomes all the difficulties encountered by surgeons viz, the difficulties in
12

using haemostatic agents and plain antiseptic solutions, as well as an anti-microbial agents separately.
Another method of use is saturating a guaze strip with 1 to 5% w/v of the Feracrylum in
water or physiological solution and then drying the gauze strip for later use.
As evidenced clinically, the solution can be repeatedly sterilized with no decomposition and retains its anti-microbial properties.
The present invention is described in more detailed below by examples which are illustrations but not restrictive.
Following are the examples of the descriptive process for manufacturing of claimed items which are not limiting improvements / modifications in the preparation of the same is obvious to one skilled in the art of formulation.
Example 1
10 litres of Feracrylum solution prepared as per the process comprising of adding of Acrylic acid to initiator in the molar ratio of 166:186:1 and Acrylic acid to iron salt in molar ratio of 105:115:1. The same solution is fed into a feed tank continuously with the hot air kept at 140°C and is passed through the drying chamber, evaporated by the hot air and the powder particles are separated from the water vapour which is filtered through bag filter. The outlet temperature is maintained at 85°C. The closed system maintains the sterility of the product without cross contamination and the powder is
collected through cyclone separator. Spray dried Feracrylum with particle size less than 100 μ is received and packed in a tight container. The pharmaceutically acceptable
Feracrylum powder so received meets with all the quality specification as specified above on page 6.
Example 2
In a glass vessel, which has arrangement for stirring and temperature control, 800 ml of double distilled water is taken, temperature maintained at 25 °C in which 10 gms of Feracrylum (spray dried) is slowly added under continuous stirring till all the material is dissolved. The mixture is made to 1 litre and then filtered aseptically through a
13

0.45 μ filter. The pH of the solution is measured potentiometrically and maintained between 2.5 to 4.5. The solution is filled in a suitable container and further terminal sterilization is carried out as is conventionally undertaken for sterile preparations. The same is filled in 1ml to 50 ml of FFS bottle for local application.
Example 3
In a glass vessel, which has arrangement for stirring and temperature control, 800 ml of double distilled water is taken temperature maintained at 25°C in which 120 gms of Feracrylum (spray dried ) is slowly added under continuous stirring till all the material is dissolved. The mixture is made to 1 litre and then filtered through a 0.45 a, filter. The pH of the solution is measured potentiometrically and maintained between 2.5 to 4.5. The solution is filled in a suitable container and further terminal sterilization is carried out as is conventionally undertaken for sterile preparations or sterilize by y sterilization. This can be used as lotion after filling in bottle of 2 - 5ml packing with accessories like ear buds for use by dipping and applying to the bleeding cut surfaces.
Example 4
3% aqueous solution that may obtain as example II is filled in 15 ml glass bottle with crimped ferrule containing visible drip tube with pumping and Spraying gadget for use as Spray for wounds caused due to injury of voluntarily and involuntarily wounds.
Example 5
For preparing 3% Feracrylum gel, 3% Feracrylum is taken with 5% Carbopol 934 under constant stirring. Cool and add triethanolamine to pH 7. Add 12.5% of Propylene glycol and 0.5% of Butylated Hydroxy Anisole. The total weight of the product is made to 100 gms using water as a vehicle. Cool and set the gel for filling in 5 to 200gm tubes for local application as wound healing and for clinical application in diabetic foot ulcers.
Example 6
100 gm of PEG 40 and 4000 are taken in molar ratio and 6 gms of Spray dried Feracrylum is sprinkled uniformly and mass of PEG is molten and cooled and
14

spreaded uniformly on cotton/rayon gauze. Cut in 10 x 10 cm size in single or multiple pack and sterilize with y radiation. The usefulness of such tulles are for surgical wounds, post operative surgery etc.
The above formulations can be modified with addition of the same with 1% Framycetin or 1% Neomycin or 1% Clotrimazole for application on wounds of bacterial and fungal infections.
Example 7
Further in a separate formulations example 5 can be modified by mixing 1% Curcumin
or other natural antibacterials for application on infected wounds.
Example 8
3% gel can be prepared further for painful condition of wounds by mixing the
composition of example 5 with 1% Aceclofenac or 1% Diclofenac sodium or other
NSAID for painful conditions after orthopaedic surgery.
Example 9
1% to 3% of Bentonite is mixed with 3% - 5% of Spray dried Feracrylum and Sprayed on cotton and uniformly spread, on roller bandage and then used after hydration for the dressings of post operative orthopaedic surgery.
ExamplelO
300 mg Feracrylum capsules of Spray dried Feracrylum is prepared by mixing 30.77% of Spray dried Feracrylum, 60.0% lactose, 1.56% aerosil and 7.69% talcum. Mix thoroughly and fill in gelatin capsules for treatment of//, pylori and dyespesia.
Example 11
Further the same capsule as cited in example is mixed with 20 mg of H2 antagonist like Omeoprazole or 20mg to 40 mg Lansoprazole, Rabeprazole, Pantoprazole for the treatment of dyspepsia induced or in oesophagal refluxes or for treating of H, pylori
Example 12
Take 3% Feracrylum in a vessel containing water. Add 5% carbopol 934 with constant stirring. Adjust the pH with triethanolamine upto pH 7. Add 0.5% of Centbucridine

15
Hydrochloride previously dissolved in water and 15% Dimethyl Isosorbide. Add 10% of Propylene glycol, stir well and make it to 400 gms jusing water. Continuously stir and adjust the pH to 5 to 5.5. Allow to set the gel and fill in 5 to 50 gm tubes for the local application as an antipruritic and bleeding haemorroids
Example 13
The prepared gel as given above is mixed with 0.5% to 0.1% Lignocaine hydrochloride for the application in bleeding piles.
Example 14
The 100 mg tablets of Feracrylum is prepared by taking 15.621% Spray dried Feracrylum mixed with 16.0% starch, 32% dicalcium phosphate, 29% microcrystalline cellulose, 3% starch., 0.8 % gelatin, 0.4% magnesium stearate and mixed with disulegrate like 0.8% pregelatinized starch. The tablets are compressed and packed. The same can be used for oral formulation for treating H. pylori and bleeding ulcers.
Example 15
The composition with the Spray dried Feracrylum of present invention was studied to ensure their efficacy in various forms of the composition. The synergism between the excipient and the active in the syrup, spray in terms of efficacy is demonstrated as
follows.

Description Placebo Spray Spray prepared by US 4,215,106 Spray prepared by the present process
Composition Nil 3% Feracrylum 3% Feracrylum
Excipient Water Water Water
Hydrofluoroalkane 10g 10g 10g
Both the above samples were compared in terms of efficacy and synergism with the excipients. It was observed that the Feracrylum spray prepared by Spray dried powder has a better in-vitro antimicrobial activity as compared to placebo and spray prepared using prior art knowledge of Feracrylum.

Determination of static of sprays :
I. Procedure for determination of Antibacterial static activity of sprays.
The spray was actuated in the laminar flow unit and the sample was collected in a
sterile conical flask.
Foam was allowed to subside.
With a sterile pipette calculated amounts of the sample solution were pipette out in
sterile test tubes.
Volume was made up to 10 ml using sterile broth recommended for the particular organism under test. Loopful of standardized culture of the organism being tested, was inoculated in each test tube except for the negative control.
Test tubes were incubated at 37°C for 24 hrs. Turbidity produced was observed and compared with positive control.
The minimum concentration of the sample which showed very little turbidity due to inhibition of growth of the innoculated culture was concluded to be MIC.
Activity was tested against following organisms :
i Candida albicans
ii Pseudomonas aeruginosa
Standardization of the innoculum :
This was carried out using 24 hrs. old culture and transferring the growth in saline. The optical density of this innoculum was then adjusted to 0.3 O.D. using photoelectric colorimeter 112, filter no. 8, and range above 360 nm.
II Procedure for determination of cidal activity of sprays :
Initial procedure followed was same as for static activity determination.
Test tubes were incubated at 37°C for 24 hrs. after innoculation.
17

Loopful of sample from each test tube was streaked on agar medium recommended for the organism under test. Plates were then incubated at 37°C for another 24 hrs. and observation for growth produced was done.
The minimum concentration of the sample which did not show any growth was concluded to be the Minimum Bactericidal Concentration (MBC).
3% spray of non spray and Spray dried Feracrylum
Static/cidal activity of the materials were observed, as follows:. Comparative Antimicrobial Activity

Name of the Organism Spray Dried 3% FeracrylumSprayInhibition ConcentrationsStatic Cidal 3% Feracrylum by Non Spray dried Feracrylum Inhibition Concnetrations Static Cidal
Pseudomonas aeruginosa 0.3% 0.3% 1.0% 1.5%
Staphyloccous Aureus 0.2% 0.4% 2.2% 2.6%
Candida albicans 0.4% 0.4% 2.9% 3.75%
Conclusion : From the above in-vivo experimental data it can be observed that Spray
dried Feracrylum shows better activity as well as synergism of static/cidal activity vis a
vis Feracrylum prepared by Non Spray dried Feracrylum.
Example 15:
Composition of Feracrylum syrup (Spray dried & Non Spray dried) was prepared
as under:
Feracrylum 3%
Sorbitol 10%
Sodium Saccharine 0.04%
Sun set yellow 0.002%
Sweet Orange 0.2%
Sodium Citrate 0.6%
D.M. Water (q.s. base) 100%
pH 4.52
MIC and MBC test of Feracrylum Syrup against Pseudomonas aeruginosa.

Media used : Nutrient Broth and Agar
Prodcedure:
MIC of Feracrylum was determined by the Tube Dilution Method. Concentrations of
drug Feracrylum was 30mg/ml. Serial dilutions were prepared in Nutrient Media to
obtain 0.1, 0.2, 0.3, 0.4, 0.5 , 1.0% of Feracrylum. Culture suspension of test
organism was prepared in sterile saline from 18-24 hour old slant culture. The said suspension was adjusted to 107 cells/ml to an Optical Density of 0.1 at 570 nm. 0.1 ml of this inoculum was added to each tube containing 5 ml of various dilutions of the test compound.
All the tubes were incubated at 37°C for 48 hours and results were recorded. Cidal effects were confirmed by absence of growth following streaking on solid media. The minimum concentration at which there was no growth on the plates was considered as MBC (Minimal Bactericidal Concentration) of the antibacterial agent. Appropriate controls and sets of tests were used in the testing.

Results Clarity Antimicrobial activity - Clear Yellow Solution
Test Organisms Concentration of Spray dried Feracrylum Static activity Cidal activityof Spray dried Cidal activity Concentration of Non Spray dried FeracrylumStatic activity Cidal activity
Pseudomonas aeruginosa 0.20% 0.50% 1.5% 2.5%
Conclusion :
From the above result it is very clear that there exist a synergism between the excipient and the Spray dried Feracrylum in terms of better in-vitro antibacterial activity vis a vis syrup prepared from Non Spray dried Feracrylum. This can be attributed to better dispersion in terms of particle and the method for the preparation of Feracrylum.
Example 16
35.708% of Spray dried Feracylum is mixed with 14.6% starch, 24.6% dicalcium phosphate, 22% microcrystalline cellulose, 0.64% gelatin, 0.309 magnesium stearate
19

and mixed with disulegrate like 0.0119% aerosil, 0.619% pregelatinized starch and 1.226% acidisol. These tablets are compressed to give 300 mg Feracrylum for the application in treating H.pylori infections by taking orally. The results of the Drug dissolution by so prepared formulation are compared with Non Spray dried Feracrylum.
FERACRYLUM DISSOLUTION STUDY
Apparatus : USP XXVIII Type I Medium : Water
Temp : 37°C
% Feracrylum Dissolution of Spray dried Feracrylum vs. Non Spray dried Feracrylum

Tablet Formulation Capsule Formulation
Time 15' 30' 15' 30'
Non Spray Dried 30% 45% 25% 40%
Spray Dried 60% 98% 67% 99%
The above results of Feracrylum dissolution indicates that formulation prepared with Spray dried powder has better in-vitro dissolution characteristic when compared against non Spray dried preparation of capsules can be co-related with the in vivo wound healing activity.
From the above examples it may be derived that Spray dried powder of Feracrylum prepared with above test, quality and with particle size of max. l00μ has better dispersibility, spreadiblity and availability in terms of in vitro and in vivo availability.
Example 16 : Wound Healing Activity of prepared Feracrylum Topical Gels
Topical gels were developed by taking 3% w/w Feracrylum using different gelling agents like 5% Carbopol 934, triethanolamine, 12.5% of Propylene glycol and 0.5% of Butylated Hydroxy Anisole. The gels were prepared the total weight of the product was made to 100 gms using water as a vehicle.
Male wistar rats weighing 150-220 g were used after approval of the institutional ethics committee. They were kept on standard pellet diet. The total of five groups each having siz animals were used. The excision of wound was created by cutting away circular

20
piece (20 mm2) of skin in its full thickness from interscapular region to monitor wound contraction and period of epithelisation. Sufficient amount of formulations (approximately 2 g) were applied to cover the wound. The wound contraction was
calculated as % reduction in wound area. The progressive changes in the wound area were monitored planimetrically by periodically (every alternate day) tracing the wound margin on a transparent paper. The days required for falling of eschar leaving no raw wound was taken as period of epithelisation. The data obtained was subjected to statistically analysis (ANOVA) using software "INSTANT" by Tukey -Karmar multiple comparison test. The marketed silver sulfadiazine was used as standard. Topical antimicrobial plays an important role in preventing contamination of wound and entry of microorganism into wound. This will accelerate healing process of wound. The results of wound healing activity are shown in Table I and presented in Figure 1. Topical gel of 3% w/w Spray dried Feracrylum prepared showed significant wound healing activity as compared to control and standard and 3% gel prepared with Non Sprayed material.
From the overall results it can be concluded that Feracrylum in concentration of 3% w/w as topical gel prepared with Spray dried powder of Feracrylum was having good wound healing activity.
Table I: Closure of Excision Wound Area on Post Wounding Days

Wound Area (m Post Wounding days m2 ± SD) Control (Groupl) Spray dried FeracrylumGel(Group II) Non Spray dried Feracrylum Gel (Group III)
0 20.02±0,81 (0%) 20.06±0.78 (0%) 19.04±0.71 (0%)
2 19.03±1.13 (6.6%) 13.39±0.53 (33.2%) 17.04±0.69 (10.5%)
4 17.05±0.93 (14.8%) 12.37±0.70*** (38.3%) 15.03±0.64 (21.0%)
6 16.04±0.70 (!9.8%) 6.72±0.62*** (66.5%) 12.04±0.81* (36.7%)
8 15.06±1.04 (24.7%) 4.39±0.46*** (78.1%) 10.04±0.66* (47.2%)
10 13.04±0.81 (34.8%) 3.37±0.45*** 83.5% 7.02±0.72** 63.1%)
12 11.04±0.90 (44.8%) 0.38±0.05*** (98.1%) 4.72±0.60*** (75.2%)
21
Values are mean ±SD of six animals (n=6) each group. Numbers in parenthesis
indicates % Wound Contraction.
Values are significant at ***p<0.001, **p<0.01, P<0.01,P<0.05.
Example 17: To study the open wound healing activity of the Tulle prepared from Spray dried Feracrylum in Sprague Dawley Rats
Compound: Feracrylum Spray Dried 3% Tulle Control: Feracrylum Non Spray Dried 3% Tulle
Animals: Sprague Dawley rats (n=6) weight 350-450g of either sex bred in our animal colony from the original stock obtained from National Institute of Nutrition were used for the study.
Housing Conditions: The animals were housed 3 per cage in polypropylene cages and were moved to experimental from where they were allowed to acclimatize for a day before treatment. The room environmental conditions had design specifications as follows:
10% air exhaust in the air conditioning unit Relative humidity of 60 ± 5%
Temperature 25 ± 3° C
12 hour light/dark cycle
Amrut certified rodent diet (Maharashtra Chakan Oil Mills Ltd.) and tap water (boiling
hot water colled to room temperature), provided ad libitum.
Treatment:
One day before the study animals were removed from their cages and were put in a holding bucket for randomization and were divided into groups and numbered. After allocation of the numbers to animals, the cages were given respective identification numbers.
On the day of the study, the animals were removed from the cages and four circular areas of approximately 100 sq. mm were marked out with India Ink on either side of the
22

inter scapular region and it's full thickness were excised with a scalpel and scissors under ether anaesthesia.
3% Feracrylum tulle prepared using 3% Feracrylum Spray Dried powder and Non Spray Dried powder and tulle was applied over incised wound and covered with above
tulles for treating the wound. OBSERVATIONS:

Observations Date of Study Groups
Wound Contraction Before start of treatment Every 4 days All
Mortality / Moribund Daily All
Data Evaluation End of treatment All
Preparation of report End of study
DATA EVALUATION:
The progressive decrease in the wound are were monitored every four days tracing the
wound margin on a tracing paper and the area were assessed using a graph paper. The
wound contractions were measured as a percentage reduction in the wound area as
depicted in table below.
RESULTS:
Table II : Excision wound studies showing percentage reduction in wound size when treated with the test compound

Group Day 4 Day 8 Day 12 Day 16
TestFeracrylum Spray Dried 3% Tulle 63.90 ±5.20 77.32 ±4.34 100.00 ±0.00 100.00 ±0.00
Control Feracrylum Non Spray ried 3% Tulle 50.96 ± 2.96 62.96 ± 6.73 88.11 ± 4.45 100.00 ±0.00
Values are expressed as Mean ± SEM
Conclusion :
Test compound showed a good wound healing property in comparison to the control.
23

Example 18: In-vitro efficacy studies of Spray dried Feracrylum powder and the formulation of tablet prepared thereof.
Peptic Ulcers;
Peptic ulcers are chronic, most often solitary, lesions that occur in any portion of the gastrointestinal tract exposed to the aggressive action of acid-peptic juices. By definition they are defects in the mucosa that penetrate atleast into the submucosa, and usually into the muscularis propria or deeper. Most are round, sharply punched out craters 2-4cm in diameter. Those in the duodenum tend to be smaller, and occasional gastric lesions are significantly larger.
The major forms of peptic ulcer are duodenal ulcer (DU) and gastric ulcer (GU) both of which are chronic diseases often caused by the bacterium Helicobacter pylori.
The term Peptic ulcer also encompasses gastric ulcers and duodenal ulcers associated with stress or the ingestion of drugs most commonly aspirin and other NSAIDs. Peptic ulcer develops when gastroduodenal mucosal defences are unable to protect the epithelium from the corrosive effects of acid and pepsin.
Sites:
Favoured sites are the anterior and posterior walls of the first portion of the duodenum and the lesser curvature of the stomach.
Pathogenesis:
There are two key facts.
1) The fundamental requisite for peptic ulceration is mucosal exposure to gastric acid and pepsin.
2) There is a very strong causal association with Helicobacter pylori infection.
Other Factors which Cause Peptic Ulcers:
Chronic use of NSAIDs and Aspirin suppresses prostaglandin synthesis. Cigarette smoking impairs mucosal blood flow and healing.
24

Corticosteroids in high dose and with repeated use promote ulcer formation.
Clinical Features of Peptic Ulcers:
Most Peptic ulcers caused epigastric gnawing, burning or boring pain, but a significant minority, first come to light with complications such as haemorrhage or perforation. The pain tends to be worse at night and occurs usually 1-3 hours after meals during the day. It is usually relieved within a few minutes by food or antacids, presumably as a result of partial neutralization of gastric acid.
Nausea, vomiting, bloating, belching and significant weight loss are additional manifestation. Bleeding is the chief complication, occuring in upto 1/3 of patients and may be life threatening.
Malignant transformation is unknown with duodenal ulcer and is very rare with gastric ulcers.
Peptic ulcers are notoriously chronic, recurrent lesions which more often impair the quality of life then shorten it when untreated, the average individual requires 15 years for healing of a peptic ulcer.
Diagnosis:
For initial diagnosis and endoscopy should be performed to look for any lesion in the stomach and duodenum and for taking biopsy specimens for the urease test and/or histological examination.
Helicobacter pylori can be identified by histologic examination, culture, and detection of urease activity. On stained tissue sections, Helicobacter pylori is positive for Giemsa and Warthin-starry stains and faintly positive for Hematoxylin. The organisms can be cultured successfully from gastric secretions. Helicobacter pylori produces large amounts of urease. The rapid urease test of gastric biopsy material is relatively simple and reliable method for identification of Helicobacter pylori. A positive test results from an increase in pH, with the phenol red indicator turning from light orange
25

to red. The test is inexpensive, with a sensitivity of atleast 90% and specificity approaching 100%.
A urea breath test using 13C or 14C also has been developed for Helicobacter pylori on the basis of urease production with release of carbondioxide. This test has a sensitivity of 90-95% and a specificity of 98-99%.
More non-invasive tests are being developed like stool antigen for Helicobacter pylori -HpSA, PCR, CagA region estimation etc,
Antibodies (IgG and IgA) to Helicobacter pylori has been identified in sera of individuals with Helicobacter pylori colonization. Serology is now used widely for routine detection of Helicobacter pylori. An enzyme linked immunosorbent assay (ELIS A) in the most commonly used serologic method, and a variety of diagnostic kits are available commercially.

Diagnostic Tests for Helicobacter pylori
Test Sensitivity (%) Comments
Urease assay Breath Biopsy 90-95 90-98 Simple; may be used to monitor therapy Requires endoscopy
Histology 70-99 Requires endoscopy; may require special Stains
Culture 70-95 Requires endoscopy; may be essential if antibiotic resistance emerges
Serology 95 Does not differentiate between active and remote infection; epidemiologic tool; titer decreases slowly after eradication
Treatment:
Helicobacter pylori eradication therapy has been shown to reduce ulcer recurrence over 6-12 month period following initial ulcer healing, ulcer recurrence rate have been demonstrated to be 85% in Helicobacter pylori positive patients but only 10% in Helicobacter pylori negative patients. Thus Helicobacter pylori eradication therapy is recommended for all Helicobacter pylori positive patients with confirmed gastric or duodenal ulcers both in patients presenting with first ulcer and those presenting with recurrence.
26
In 1994, WHO recognised Helicobacter pylori as a grade I (definite) carcinogen and the NIH consensus development conference statement recommended that all patients with duodenal ulcer or gastric ulcer and concurrent Helicobacter pylori infection should receive treatment aimed at eradicating the bacteria.
In 1997, Asia-Pacific Consensus Conference and European Consensus panel strongly recommended that the following patients with Helicobacter pylori infection should be treated :
1) Peptic ulcer.
2) Low grade MALT lymphoma.
3) Severe macroscopic or microscopic gastritis.
4) Recently resected early gastric cancer.
In addition treatment aimed at eradication was advisable in Helicobacter pylori-infected individuals with:
a) Non-ulcer dyspepsia.
b) Family history of gastric cancer.
c) GERE) necessitating long term PPI therapy.
d) Planned or ongoing NSAID ingestion.
e) Following gastric surgery for peptic ulcer disease or gastric cancer.
In the Indian context reinfection rate is high; hence eradication regime for patients with complications is advisable. Relapse rates following successful eradication of Helicobacter pylori has been reported to be lower (less than 2 present) compared to that following acid suppressant therapy alone (80%).

Therapeutic Regimens
Antibiotic + PPI or RBC Eradication rate Duration
1) Dual therapy
a) Omeprazole + Amoxycillin 50-85% 2 weeks
b) Omeprazole + Clarithromycin 80% 2 weeks
2) Triple therapya) Classical triple therapy
Bismuth + Metronidazole + Tetracycline b) Alternative triple therapy 60-90% 2 weeks
Omeprazole + Amoxycillin + Tinidazole 95% 1 week
Omeprazole + Amoxycillin + Clarithromycin 90% 1 week
3) Quadruple therapyTwo antibiotics + Two adjunctives
27

a) Omeprazole + CBS + Tetracycline +Metronidazoleb) Omeprazole + CBS + Amoxycillin +Secnidazole 85-95% 85-95% 1 week 1 week
Definition of Cure:
This may be defined as absence of the organism by tests performed no sooner than 4 weeks after cessation of antimicrobial therapy.
Nevertheless, with present day therapies (including antibiotics active against Helicobacter pylori), patients get palliative treatment or treatment for regression of ulcers and are thus saved from the surgeons knife.
The present inventors, for the first time have carried out application of Feracrylum in the form of oral tablet formulation for eradication and treatment of Dyspepsia associated with H.pylori infection in peptic and duodenal ulcers.
The formulation of Feracrylum is been termed as Spray dried Feracrylum and following tests are carried out for the compound Spray dried Feracrylum.
1) Summary of the in vitro results
As it may be known, there are several pathogenic strains of Helicobacter pylori which are isolated and characterized (viz., Genotypes) and the cure of such virulent strains poses a challenge to the medical profession. Further vis-a-vis these enteric pathogens, specifically Helicobacter pylori, have the risk of developing gastric adeno carcinoma and Mucosa Associated Lymphoid Tissue (MALT) lymphoma.
About 24 strains of various genotypes have been tested for Spray dried Feracrylum. Data generated through studies carried out at an internationally acclaimed institute has revealed that Spray dried Feracrylum, when tested in vitro, is active against Helicobacter pylori pathogenic strains.
The study protocol included the test against isolated strains (genetically tagged) and species of Helicobacter pylori.
28

These strains were isolated from patients manifesting mostly Duodenal Ulcer, one case of Gastric malignancy, one case of Duodenitis and three cases of post therapy healed
ulcer.
Method
Agar dilution method was used to determine the minimal concentration of Spray dried Feracrylum required to inhibit the growth of Helicobacter pylori. The drug was dissolved in minimum volume of sterile distilled water to get different Spray dried Feracrylum concentrations used in this drug sensitivity study. In this case the following drug concentrations were used: 0.2 mg/ml, 0.4 mg/ml, 1.0 mg/ml, 2.0 mg/ml, 3.3 mg/ml and 6.6 mg/ml.
A total of 24 different H.pylori strains were selected for this study. Blood agar plates were prepared with brain-heart infusion agar containing 10% defibrinated sheep blood and a particular dilution of the drug added to each plate. Control plates with no Spray
dried Feracrylum were simultaneously prepared. The drug-supplemented as well as the control blood-agar plates were streak-inoculated with ~ 104 CFU of each H.pylori
strain. The inoculated plates were incubated at 37°C under 10% CO2 and 5% O2 and were observed for the presence of bacterial growth initially at 48 hours and then daily for up to 10 days.
Results:
With the first set of 8 H.pylori strains, all the drug concentration i.e., from 0.2 mg/ml to 6.6 mg/ml were tested. It was observed that using the drug concentrations of 0.2 mg/ml to 2.0 mg/ml all the 8 strains had grown well in all the sectors. However, with 3.3 mg.ml drug, growth was observed in 4 of the 8 sectors while 6.6 mg/ml there was no growth in any sector.
Subsequently with all H.pylori strains, the following three concentrations of Spray dried Feracrylum were tested - 3.3 mg/ml, 4.0 mg/ml and 6.6 mg/ml. All the results indicated that 4.0 mg/ml Spray dried Feracrylum could be considered the lowest diluted drug concentration that prevented visible bacterial growth.
The list of the strains, their respective genotype and their sensitivity to Spray dried Feracrylum are shown in Table 1 and 2.
29

TABLE- 1
H.pylori strains used : 109A, 114A, 100A, 86A, 115B, 91 A, 62A, 77A, 222B, 278A,
286B, 291B, 105A, 331A, 353A, 324B, 41B, 56B, 60A, 17A, 21B, 78B, 111A and 27B.
Medium used : Brain-heart infusion agar + 10% defibrinated sheep blood
Volume of : 25 ml
medium/plate

TCL FA-99 added/ml Growth of strains observed after 7 days from plating
109 A 114 A 100 A 86 A 115 B 91 A 62 A 77 A 222 B 278 A 286 B 291 B 105 A 331 A 353 A 324 B 41 B 56 B 60A 17 A 21 B 78 B Ill A 27 B
3.3 mg/ml - + - - - + - + + - + + - + + + + + + + - - - +
4.0 mg/ml - - - - - - - - - - - - - - - - - - - - - - -
6.6 mg/ml - - - - - - - - - - - - - - - - - - - - - - - -
30

TABLE - 2

Sr.N o. Strain No. Clinical manifestation of patient from whom the strain was isolated Genotype of strains
Vac A mlb Vac A m2 Vac A SI Vac As2 CagA Rt. EndOfcag PAI iceAl iceA2
1. 109 A D.U. - + + - + Type III + -
2. 114A D.U. + - + - + Type III + -
3. 100 A Gastric malignancy - - + - + Type III - +
4. 86A D.U. - + + - + Type III + +
5. 115B D.U. + - + - + Type III - +
6. 91A D.U. + - + - + Type III + -
7. 62A D.U. - + + - + Type III + -
8. 77A D.U. + - + - + Type III - +
9. 222B Duodenitis - + - - + - +
,0. 278B Ulcer healed (post therapy) + + + - + Type HI - +
11. 286B Ulcer healed (post therapy) - + + - + Type III + +
12. 291B Ulcer healed (post therapy) + + + - + Type III + -
13. 105 A D.U. + - + - + Type III + -
14. 331A D.U. - + + - + + -
15. 353A* NUD
16. 324B D.U. + - + - + + +
17. 41B D.U. + - + - + Type III + -
18. 56B D.U. - + + - + Type III - +
19. 60A D.U. + - + - + Type III + -
31

20. 17A D.U. + + + - + Type III + -
21. 21B Gastric malignancy + - + - + Type III - +
22. 78B D.U. + . + - + Type III - -
23. 111A D.U. + - + - + Type III + -
24. 27B D.U. + + + - + Type III - +
* Genotype not yet determined D.U. = Duodenal ulcer. N.U.D. = Non-ulcer dyspepsia
32

3) Summary of the clinical data
On the basis of the animal studies that had been submitted for the Compound
'SPRAY DRIED FERACRYLUM' the Drug Authorities were requested
permission to conduct Pilot Studies.
The two main indications selected were:
1. Non-ulcer Dyspepsia infected with H.Pylori
2. Bleeding Gastric & Duodenal Ulcers with or without H.Pylori
The clinical studies were carried out using Spray dried Feracrylum (300 mg tablet of Feracrylum prepared using Spray dried Feracrylum of the above specification).
The first 10 pilot cases carried out revealed that 'Spray dried Feracrylum' effectively eradicates H.Pylori and treats bleeding Duodenal Ulcers.
On the basis of the Pilot Studies, the Permission was received from the Drug Regulatory Authorities to conduct Multicenteric Clinical Trials.
The Clinical Trials commenced at the end of 2000 for which the following was undertaken-
a. Four (4) renowned Centers in the country were selected;
b. Four (4) Gastroenterologists as Investigators were selected who were each
supposed to recruit twenty (20) patients.
c. A Protocol was designed - which was checked, modified and then submitted
to the Drug Regulatory Authorities for final approval.
Following are the main features of the Protocol:
1. Patients are to be subjected to endoscopic diagnosis. Biopsies are to be taken from fundus and antrum and tested for the presence of H.Pylori. Only cases with H.Pylori Positive in both biopsies are to be included.
33

2. Rapid Urea Test (RUT) has to be conducted from both the places (fundus and
antrum).
3. simultaneously Feracrylum and Proton Pump Inhibitors (PPIs') or
H2Antagonists are to be administered during the course of study to other group for comparison , which is to last for Six (6) weeks viz., Two (2) weeks of Active treatment and Four (4) weeks of Placebo.
4. Endoscopy is to be done at Day Zero (0) and on Day Forty Two (42) viz., at the end of the Six (6) weeks.
5. Clinical evaluation, Biopsies and Rapid Urea Test (RUT) are to be done at the end of Six (6) weeks to evaluate response.
Findings from the Investigators have revealed encouraging results.
The Summary of the Results is attached herewith titled "Latest Clinical Status Report
Table 3
CLINICAL REPORT

CENTRE TOTAL NUMBER OF CASES RECRUITED RESULTS Feracrylum tablet (300 mg) Feracrylum with H2 AntagonistPantaprzole (20 mg)
No-cure Cure No-cure Cure
I 14 6 6 6 8
II 20 1 18 - 20
III 20 9 9 1 19
IV 20 2 18 1 19
74 18 51 8 66
Conclusion:
The above clinical results indicates the cure rate of H.pylori infected patients to be 69%
whereas, treatment of Feacrylum with H2 antagonist shows the cure rate of the patient at 89%.
34

Example 18 Determination of Minimum inhibitory concentration (MIC) of Spray dried Feracrylum against different enteropathogens
I) Medium : Mueller Hinton Agar
Volume of medium/ plate : 25 ml

Concentration of Strain No.
Spray dried
Feracrylum (µg/ml)
AH0 82 S.t2 EPEC 3748 EPEC A-414, LT AH0 68 VP309
128 + + + + + +
64 + + + + + +
32 + + + + + +
16 + + + + + +
8 + + + + + +
4 + + + + + +
2 + + + + + +
1 + + + + + +
0.5 + + + + + +
0.25 + + + + + +
Control + + + + + +
+ = Growth, AHO 82 = Vibrio cholerae Ol Ogawa, S.t 2 = Salmonela typhimurium, EPEC 3748 = Enteropathogenic Esherichia coli (02:H25), ETEC A-414, (LT) = Enterotoxigenic Esherichia coli (ONT:HNT), AHO 68 = Vibrio cholerae 0139, VP 309 = Vibrio parahemolyticus (08: KUT).
II)
Vibrio cholerae strains used : 181, 175, 272, 285, 219, 186, 211
(01, Ogawa, El Tor biotype)
Medium used : Mueller Hinton Agar
Volume of medium/plate : 25 ml

Spray dried Feracrylum added/ml Growth of strains observed after 18 hours from plating
35

181 175 272 285 219 186 211
3.3 mg/ml - - - - - - -
4.0 mg/ml - - - - - - -
6.6 mg/ml - - - - - - -
Control Plate + + + + + + +
It is thus evident that spray dried feracrylunm at dose range of above 3 is effecting in inhibiting the growth of Vibrio strains while a lower dose is not effective.
Example 19: For the anti microbial activity against Giardia and Entamoeba infections the following Concentration of drugs were used:

Sample no. Final concentration of Drug Spray dried Feracrylum
1A 4 mg/ml
IB 4 mg/ml
2A 5 mg/ml
2B 5 mg/ml
+CA Positive control
(Metronidazole 120 ul per tube of concentration 1 mg/ml.)
+CB Positive control
(Metronidazole 120 ul per tube of concentration 1 mg/ml.)
-CA Negative control (No drug)
-CB Negative control (No drug)
Table 4
Antimicrobial activity of Spray dried Feracrylum against Entamoeba histolytica

Sample Entamoeba histolytica
No.
15 min 30 min 45 min lhr 2hrs 3 hrs 4 hrs 5 hrs 6 hrs 24 hrs
1A + + + + - - - - - -
(50%) (80%) (90%) (100%) (100%) (100%)
IB + + + + - - - - - -
(50%) (80%) (90%) (100%) (100%) (100%)
2A + + + + - - - - - -
(60%) (90%) (100%) (100%) (100%) (100%)
2B + + + + - - - - - -
(60%) (90%) (100%) (100%) (100%) (100%)
+CA - - - - - - - - - -
(75%) (100%) (100%) (100%) (100%) (100%) (100%) (100%) (100%) (100%)
+CB - - - - - - - - - -
(75%) (100%) (100%) (100%) (100%) (100%) (100%) (100%) (100%) (100%)
-CA + + + + + + + + + +
-CB + + + + + + + + + +
36

+ Live and attached cells.
- (%) Dead and unattached cells, (percentage of d
Table 5
Antimicrobial activity of Spray dried Feracrylum against Giardia lamblia

Sample No. Giardia lamblia
15 min 30 min 45 min 1 hr 2hrs 3 hrs 4 hrs 5 hrs 6 hrs 24 hrs
1A + + + + + + + + + -
IB + + + + + + + + + (100%)
2A + + + + + + + + + (100%)
2B + + + + + + + + + (100%)
+CA + + + + + + (100%)
+CB + + + + + + (80%) (100%) (100%) (100%)
-CA + + + + + + (80%)+ (100%)+ (100%)+ (100%)+
-CB + + + + + + + + + +
+ Live and attached cells.
- (%) Dead and unattached cells, (percentage of death).
So, The drug Spray dried Feracrylum is active in vitro against axenically cultured Giardia lamblia (Protland I ) and Entamoeba histolytica (HM1) at concentrations 4 mg/ml - 24 hrs and 4 mg/ml - 2 hrs respectively

37
We Claim :
1) A process for preparation of pharmaceutically acceptable Feracrylum powder
by Spray drying such that the feracrylum powder produced is of fine dispersibility with acceptable quality parameters and a particle size distribution of below l00µ, said process comprising: providing feracrylum solution;
supplying air through micron filters and heating at 135°C to 145°C; spraying Feracrylum in a drying chamber kept at 140°C and vacuum is maintained at -20 to -30 mm WC;
collecting Feracrylum in bag filter at a filter pressure of 5 - 10 mm; maintaining nozzle air pressure for atomization at 1,5 to 2.5kg/cm2 and feed rate of the solution at 50 ml/min to 100 ml/min
2. A process as claimed in claim 1 wherein the incoming air is filtered through
prefilter followed by HEPA filter.
3. A process as claimed in claim 1 or 2 wherein the atomization is carried out with oil free compressed air.
4. A process as claimed in any preceding claim wherein the pulse jet bag filter for collection of spray dried powder is provided with Teflon lining to minimize loss.
5. Process as claimed in claim 1 wherein the feracrylum produced has particle size of 60-70µ.
6. A process as claimed in any preceding claim wherein the Spray dried Feracrylum powder is adapted to form a clear, stable, limpid solution with water.
7. A composition comprising 1 to 12% w/v Spray dried Feracrylum having particle size below l00µ, prepared by the process comprising
i) providing feracrylum solution;
ii) supplying air through micron filters and heating at 135°C to 145°C;
38

iii) spraying Feracrylum in a drying chamber kept at 140°C and
vacuum is maintained at -20 to -30 mm WC;
iv) collecting Feracrylum in bag filter at a filter pressure of 5 - 10
mm;
v) maintaining nozzle air pressure for atomization at 1.5 to 2.5 kg/cm2 and feed rate of the solution at 50 ml/min to 100 ml/min;
and pharmaceutically acceptable excipients, the composition having a pH of 2.5-6.5
8. Composition as claimed in claim 7 wherein the composition is in the form of tablet,
solution, spray, powder, ointment, cream, gel, lotion, suppository, tulles/bandages,
surgical dressing, gargles, tampons and the like for local application and for treating
wounds externally and internally.
9. Composition as claimed in claim 7 or 8 wherein the pharmaceutically acceptable
excipients are selected from fillers, compression aids, binders, flavorings, coating
agents, sugar, colourant, gelatin, starch, PVC, PVP, methyl cellulose, CMC, HPMC,
alginate and coolant like menthol, stabilizers like parabens and polymers expanding
material for better spreadability.
10. Composition as claimed in claim 7 to 9 further comprising anaesthetics, antibiotics, analgesics, antiseptics, antibacterial, antifungals and natural products like curcumin or products of Aloe barbadensis and other natural products having antibacterial, antifungal activity, antipruritic substances like diphenhydramine HC1, Calamine, various steroids alone or in combinations.
11. Composition as claimed in claim 7 to 10 wherein anaesthetics is selected from lignocaine, centbucridine, bupivaccaine, tetracaine.
12. Composition as claimed in claim 7 to 10 wherein antibiotics is selected from polymixin derivaties, soframycin, neomycin, clindamycin.
39

13. Composition as claimed in claim 7 to 10 wherein analgesics of NSAID selected from diclofenac, aceclofenac, piroxicam, tramadol, ibuprofen, indomethacin, naproxen.
14. Composition as claimed in claim 7 to 10 wherein antiseptics are selected from povidone iodine, triclosan.
15. Composition as claimed in claim 7 to 10 wherein antibacterial are selected from nitroimidazole, metronidazole, secnidazole or its derivatives
16. Composition as claimed in claim 7 to 10 wherein antifungals namely azoles or allylamines selected from clotrimazole, fluconazole, itraconazole, terbinafine,
17.Composition as claimed in claim 7 to 16 wherein the composition is haemostatic, anti-bacterial, antiprotozoal, anti-inflammatory, antifungal and for treating dyspepsia
18.Composition as claimed in claim 7 to 17 wherein the tablets are adapted for the treatment of H.pylori and giardiasis and for cholera
19.Composition of claim 7 to 18 wherein the Spray dried Feracrylum and excipient is mixed with H2 antagonists for treatment of gastric and duodenal ulcer.
Dated this 24th day of July 2006


Title: PROCESS FOR PREPARATION OF SPRAY DRIED FERACRYLUM AND COMPOSITION COMPRISING THE SAME
ABSTRACT
A process for Feracrylum powder by Spray drying such that the feracrylum powder produced is of fine dispersibility with acceptable quality parameters and a particle size distribution of below l00µ, said process comprising providing feracrylum solution; supplying air through micron filters and heating at 135°C to 145°C; spraying Feracrylum in a drying chamber kept at 140°C and vacuum is maintained at -20 to -30 mm WC; collecting Feracrylum in bag filter at a filter pressure of 5 - 10 mm; maintaining nozzle air pressure for atomization at 1.5 to 2.5kg/cm and feed rate of the solution at 50 ml/min to 100 ml/min. the invention also relates to a composition comprising 1 to 12% w/v Spray dried Feracrylum having particle size below 100a, prepared by the process comprising providing feracrylum solution; supplying air through micron filters and heating at 135°C to 145°C; spraying Feracrylum in a drying chamber kept at 140°C and vacuum is maintained at -20 to -30 mm WC; collecting Feracrylum in bag filter at a filter pressure of 5 - 10 mm;' maintaining nozzle air pressure for atomization at 1.5 to 2.5 kg/cm2 and feed rate of the solution at 50 ml/min to 100 ml/min; and pharmaceutically acceptable excipients, the composition having a pH of 2.5-6.5
41