US20180344773A1 - Camel Milk Based Pharmaceutical Composition - Google Patents
Camel Milk Based Pharmaceutical Composition Download PDFInfo
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- US20180344773A1 US20180344773A1 US15/986,159 US201815986159A US2018344773A1 US 20180344773 A1 US20180344773 A1 US 20180344773A1 US 201815986159 A US201815986159 A US 201815986159A US 2018344773 A1 US2018344773 A1 US 2018344773A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/20—Milk; Whey; Colostrum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
- A61K39/05—Actinobacteria, e.g. Actinomyces, Streptomyces, Nocardia, Bifidobacterium, Gardnerella, Corynebacterium; Propionibacterium
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/10—Anti-acne agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
Definitions
- the present invention relates to a pharmaceutical composition for the treatment or prevention of skin or mucus membrane infections such as acne, and more particularly, the invention relates to a pharmaceutical composition based on immunized camelid milk, particularly Camelus dromedarius as the active ingredient, and a process of preparing the composition.
- Propionibacterium acne is the most common cutaneous disorder with a prevalence of 70-85% in adolescents. Although acne is not a life-threatening disease, it has significant physical and psychological effects such as permanent scarring, poor self-image, social inhibition, depression, anxiety, and suicidal tendency. Therefore, acne may be regarded as a serious medical condition.
- Topical therapy is inevitable in acne treatment and is mainly indicated in the mild to moderate acne. In more severe forms, a combined topical and systemic therapy is recommended.
- the available topical agents have a direct or indirect influence on the patho-genetic factors and are selected according to the predominant type of acne lesions.
- Animal milk has been used in the preparation of pharmaceutical and cosmetic compositions. Milk of ruminants, and predominantly bovine milk, has been utilized most. Among drawbacks of cow milk is a wide-spread allergy to it, affecting in several of its forms as much as 50% individuals in some populations. It is an object of this invention to provide a composition comprising immunized milk which keeps all the benign properties of milk but is free of the drawbacks related to cow milk. Camel milk has been traditionally used by certain ethnic groups, and it was found that, in some respects, its composition is closer to the human milk than cow milk.
- Immunized milk products are preparations made of such hyper-immune colostrum or antibodies enriched from it. These preparations can be used to give effective specific protection against different diseases.
- Colostral immunoglobulin supplements designed for farm animals are commercially available in many countries.
- immunized milk products that contain specific antibodies against certain pathogens have been launched in the market. A number of clinical studies are currently in progress to evaluate the efficacy of immunized milks in the prevention and treatment of various human infections, including those caused by antibiotic resistant bacteria.
- Bovine colostrum-based immunized milk products are used as prophylaxis against various infectious diseases in humans. Immunized milk products are examples of health-promoting functional foods, or nutraceuticals.
- a second object of the present invention is to provide a process of preparing the composition comprising;
- Another object of the present invention is to provide a method for the treatment or prevention of skin or mucus membrane infections, comprising applying to infected areas of the human body an effective amount of the composition of the invention.
- the camelid described in the invention is chosen from the genus Camelus, Llama, or Vicuna . More preferably, the camelid is Camelus dromedaries.
- the camelid described in the invention is immunized with skin or mucus membrane pathogen, preferably propionobacterium acne, Staphylococcus aureus, Streptococcus mutans, Pseudomonas aeroginosa, Haemophilus influenza, Neisseriae spp. Candida spps or Chlamydia trachomatus.
- skin or mucus membrane pathogen preferably propionobacterium acne, Staphylococcus aureus, Streptococcus mutans, Pseudomonas aeroginosa, Haemophilus influenza, Neisseriae spp. Candida spps or Chlamydia trachomatus.
- the pharmaceutical composition of the invention is in the form of cream, ointment, gel, skin wash, lotion, soap, shampoo, mouth wash, vaginal wash, eye wash, tooth paste, or spray.
- FIG. 1 shows whey of immunized camel tested by Enzyme-Linked Immunosorbent Assay (ELISA) and the kinetic response over the experiment period.
- ELISA Enzyme-Linked Immunosorbent Assay
- FIG. 2 shows Coomassie SDS-PAGE gel (12.5%) for P. acne camel whey. The run was done for 30 min at 100V, then for 45 min at 150V. Load was 20 ⁇ l/well. Dilution was (3:1) with 4 ⁇ dye for all fractions.
- FIG. 3 shows bactericidal activity of whey obtained from milk of immunized camel on P. acne.
- FIG. 4 shows the growth inhibition of whey obtained from milk of immunized camel.
- FIG. 5 shows anti-acne activity of aqueous cream extract.
- the pharmaceutical composition of this invention comprises immunized camelid milk as the active ingredient in a pharmaceutically acceptable vehicle, in addition, a method for treatment or prevention of skin or mucus membrane infections is described, for example, acne vulgaris.
- the camelid preferably Arabian camel ( Camelus dromadarius ), is immunized subcutaneously with an initial dose of 3 ml of prepared skin or mucus membrane pathogen, for example, Propionibacterium acne vaccine.
- the vaccinated camel is boosted 4 times at 2 week intervals with 5 ml of vaccine for each booster.
- the Propionibacterium acne vaccine is prepared from P. acnes (NCTC 373), as shown in Examples 1 and 2.
- the pharmaceutical composition is intended for treating the skin, and may be in a pharmaceutically acceptable vehicle in the form which include, but is not limited to liquid, paste, or solid, and more particularly in the form of ointment, cream, milk, powder, impregnated pad, wipes, solution, gel, spray, suspension, lotion, shampoo, or washing base.
- the composition may also be in the form of suspension of lipid or polymer vesicle, nano sphere or micro sphere, or polymer patches and hydro-gels allowing a controlled release.
- the composition may be in anhydrous form, in aqueous form, or in the form of an emulsion.
- the components of the pharmaceutical composition and their ratios could be adjusted according to the pharmaceutically acceptable vehicle and the intended application of the pharmaceutical composition.
- the pharmaceutical composition is in the form of a cream, and in another preferred embodiment of the present invention, the pharmaceutical composition is in the form of a gel.
- the present invention further relates to a process of preparing the pharmaceutical composition, comprising:
- P. acnes were cultured on Mueller Hinton Agar (MHA) under anaerobic conditions using Gas-Pak (Oxoid) at 37° C. for 72 hrs.
- 1.5 ml of the inactivated bacteria were mixed with 1.5 ml of the Freund's adjuvant complete (SIGMA) and injected subcutaneously into a camel.
- SIGMA Freund's adjuvant complete
- the ELISA was performed.
- Flat bottomed 96 well polystyrene micro titer plates (Greiner, Germany) were coated with 100 ⁇ l of 10 ⁇ g/m Propionibacterium acne antigens in carbonate-bicarbonate buffer (pH 9.6) overnight at 4° C.
- the plates were washed 3 times with 100 ⁇ l of 0.15 M PBS (pH 7.2) containing 0.05% Tween 20 and blocked with 100 ⁇ l of 2% Bovine Serum Albumin (BSA) in PBS for one hour at Room Temperature (RT).
- BSA Bovine Serum Albumin
- the plates were washed again and 100 ⁇ l of the serum samples diluted at 1:100 in 1% BSA were added in duplicates and incubated for one hour at RT, whereas, milk samples were added in duplicates without dilution. Negative and positive control samples were incorporated in each of the plates. After washing, 100 ⁇ l of one of the horse radish peroxidase (HRP) conjugated protein A and protein G were diluted at 1:1000 in 1% BSA, and were added separately to each well. The plates were incubated for one hour at RT and washed again.
- HRP horse radish peroxidase
- Stacking gels (4%) were prepared by adding 6 ml of distilled water, 2.5 ml staking buffer (pH 6.6), 1.3 mL of 30% acrylamide-bisacrylamide solution, 100 ⁇ l of 10% Ammonium persulfate (APS), and 10 ⁇ l TEMED.
- the milk was mixed with an equal volume of sample buffer lacking B-mercaptoethanol (non-reducing conditions) (pH 6.8).
- sample buffer lacking B-mercaptoethanol (non-reducing conditions)
- electrophoresls was carried out using running buffer with pH 8.3 at 120 volts for 60-120 minutes.
- the gel was stained with Coomassie brilliant blue R-250 (except if used for immunoblot) and destained by 20% acetic acid until clear bands were seen. Results are shown in FIG. 2 .
- Antibodies isolated from whey of Camel milk immunized with killed P. acne are mixed with a standard inoculum of the bacterium and subsequently incubated for 72 hrs. The total count is determined and then compared with the total count of the standard inoculum without mixing with the antibodies. Results are shown in FIG. 3 and FIG. 4 .
- a cream formula For the treatment of acne, a cream formula has been prepared from the following components:
- Cream Formula 0.5% w/w Material weight mg/unit Camel dried whey* 15 g 2.5 Cetyl alcohol 90 g 15 Tween 20 30.9 ml 5.15 Glycerylmonostearate type1 17.1 g 2.85 MERKUR 791 60 g 10 Purified water 387 ml 64.5 Total 600 100 *Equivalent to 0.5% w/w total protein
- a gel formula For the treatment of acne, a gel formula has been prepared from the following components:
- the active ingredients of camel milk whey were re-extracted from the cream for the purpose of stability testing.
- the cream was mixed with equal volumes of PBS and incubated in water bath at 56° C. for 10 minutes. The mixture was then centrifuged at 13400 rpm for 10 minutes, using microfuge. The aqueous phase was collected and tested using ELISA. Results are shown in FIG. 5 .
- Ear thickness the infection induced rabbit showed double fold increased ear thickness compared to the normal non infected ear. In contrast, ear thickness was significantly reduced in cases of injecting pretreated bacteria with positive whey and less significantly in cases of negative whey. In addition, no significant differences were observed concerning redness, heat, formation of papules and pus.
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Abstract
Description
- The present application is a continuation of U.S. application Ser. No. 13/609,342 filed Sep. 11, 2012, titled “Camel Milk Based Pharmaceutical Composition.”
- The present invention relates to a pharmaceutical composition for the treatment or prevention of skin or mucus membrane infections such as acne, and more particularly, the invention relates to a pharmaceutical composition based on immunized camelid milk, particularly Camelus dromedarius as the active ingredient, and a process of preparing the composition.
- Acne vulgaris caused by the bacterium Propionibacterium acne (P. acne) is the most common cutaneous disorder with a prevalence of 70-85% in adolescents. Although acne is not a life-threatening disease, it has significant physical and psychological effects such as permanent scarring, poor self-image, social inhibition, depression, anxiety, and suicidal tendency. Therefore, acne may be regarded as a serious medical condition. Topical therapy is inevitable in acne treatment and is mainly indicated in the mild to moderate acne. In more severe forms, a combined topical and systemic therapy is recommended. The available topical agents have a direct or indirect influence on the patho-genetic factors and are selected according to the predominant type of acne lesions. The therapeutic success in acne and related skin disorders are highly dependent on a regular application of the topical agents over a prolonged period of time. However, disadvantages associated with the commonly used topical agents considerably affect the patient compliance and obstruct the treatment. Currently, available treatment for acne and related skin disorders is mostly based on antibiotics and retinoids. The uses of antibiotics have a lot of limitations due to development of resistance by bacteria. On the other hand, Retinoids are highly teratogenic.
- Animal milk has been used in the preparation of pharmaceutical and cosmetic compositions. Milk of ruminants, and predominantly bovine milk, has been utilized most. Among drawbacks of cow milk is a wide-spread allergy to it, affecting in several of its forms as much as 50% individuals in some populations. It is an object of this invention to provide a composition comprising immunized milk which keeps all the benign properties of milk but is free of the drawbacks related to cow milk. Camel milk has been traditionally used by certain ethnic groups, and it was found that, in some respects, its composition is closer to the human milk than cow milk.
- Passive immunity is provided to newborns by Immunoglobulins present in colostrum until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunizing a mammal with these pathogens or their antigens. Immunized milk products are preparations made of such hyper-immune colostrum or antibodies enriched from it. These preparations can be used to give effective specific protection against different diseases. Colostral immunoglobulin supplements designed for farm animals are commercially available in many countries. Also, some immunized milk products that contain specific antibodies against certain pathogens have been launched in the market. A number of clinical studies are currently in progress to evaluate the efficacy of immunized milks in the prevention and treatment of various human infections, including those caused by antibiotic resistant bacteria. Bovine colostrum-based immunized milk products are used as prophylaxis against various infectious diseases in humans. Immunized milk products are examples of health-promoting functional foods, or nutraceuticals.
- It is an object of the present invention to eliminate the disadvantages of the prior art by providing a pharmaceutical composition comprising treated immunized camelid milk in a pharmaceutically acceptable vehicle for the treatment or prevention of skin or mucus membrane infections.
- A second object of the present invention, is to provide a process of preparing the composition comprising;
-
- a. Immunizing a female camelid with a vaccine of propionobacterium acne, Staphylococcus aureus, Streptococcus mutans, Pseudomonas aeroginosa, Haemophilus influenza, Neisseriae spp. Candida spps or Chlamydia trachomatus;
- b. Obtaining milk from the camelid;
- c. Treating the milk at a temperature of 0° C. for 1-2 hours and centrifuging at 15000 rpm to remove lipids;
- d. Treating the milk with rennin or acetic acid to reduce protein content;
- e. Pasteurizing the milk at a temperature ranging from 65° C. to 72° C. for 15 minutes; and
- f. Preparing the milk in a pharmaceutically acceptable vehicle.
- Another object of the present invention is to provide a method for the treatment or prevention of skin or mucus membrane infections, comprising applying to infected areas of the human body an effective amount of the composition of the invention.
- Preferably, the camelid described in the invention is chosen from the genus Camelus, Llama, or Vicuna. More preferably, the camelid is Camelus dromedaries.
- The camelid described in the invention is immunized with skin or mucus membrane pathogen, preferably propionobacterium acne, Staphylococcus aureus, Streptococcus mutans, Pseudomonas aeroginosa, Haemophilus influenza, Neisseriae spp. Candida spps or Chlamydia trachomatus.
- Preferably, the pharmaceutical composition of the invention is in the form of cream, ointment, gel, skin wash, lotion, soap, shampoo, mouth wash, vaginal wash, eye wash, tooth paste, or spray.
-
FIG. 1 shows whey of immunized camel tested by Enzyme-Linked Immunosorbent Assay (ELISA) and the kinetic response over the experiment period. -
FIG. 2 shows Coomassie SDS-PAGE gel (12.5%) for P. acne camel whey. The run was done for 30 min at 100V, then for 45 min at 150V. Load was 20 μl/well. Dilution was (3:1) with 4× dye for all fractions. -
FIG. 3 shows bactericidal activity of whey obtained from milk of immunized camel on P. acne. -
FIG. 4 shows the growth inhibition of whey obtained from milk of immunized camel. -
FIG. 5 shows anti-acne activity of aqueous cream extract. - The pharmaceutical composition of this invention comprises immunized camelid milk as the active ingredient in a pharmaceutically acceptable vehicle, in addition, a method for treatment or prevention of skin or mucus membrane infections is described, for example, acne vulgaris.
- The camelid, preferably Arabian camel (Camelus dromadarius), is immunized subcutaneously with an initial dose of 3 ml of prepared skin or mucus membrane pathogen, for example, Propionibacterium acne vaccine. The vaccinated camel is boosted 4 times at 2 week intervals with 5 ml of vaccine for each booster.
- The Propionibacterium acne vaccine is prepared from P. acnes (NCTC 373), as shown in Examples 1 and 2.
- The pharmaceutical composition, according to the present invention, is intended for treating the skin, and may be in a pharmaceutically acceptable vehicle in the form which include, but is not limited to liquid, paste, or solid, and more particularly in the form of ointment, cream, milk, powder, impregnated pad, wipes, solution, gel, spray, suspension, lotion, shampoo, or washing base. The composition may also be in the form of suspension of lipid or polymer vesicle, nano sphere or micro sphere, or polymer patches and hydro-gels allowing a controlled release. The composition may be in anhydrous form, in aqueous form, or in the form of an emulsion. The components of the pharmaceutical composition and their ratios could be adjusted according to the pharmaceutically acceptable vehicle and the intended application of the pharmaceutical composition. In a preferred embodiment of the present invention, the pharmaceutical composition is in the form of a cream, and in another preferred embodiment of the present invention, the pharmaceutical composition is in the form of a gel.
- The present invention further relates to a process of preparing the pharmaceutical composition, comprising:
-
- 1. Immunizing a female camel with a vaccine of intact or whole cell lysate of P. acne;
- 2. Obtaining milk from the female camel;
- 3. treating the female camel milk at a temperature of 0° C. for 1-2 hours and centrifuging at 15000 rpm to remove lipids;
- 4. Treating the milk with rennin or acetic acid to reduce protein content;
- 5. Pasteurizing the camel milk at a temperature ranging from 65° C. to 72° C. for 15 minutes; and
- 6. Preparing the camel milk in a pharmaceutically acceptable vehicle.
- The invention will be further described and illustrated in the following examples.
- The following examples illustrate the present invention without, however, limiting the same thereto.
- Cell Culture
- P. acnes (NCTC 373) were cultured on Mueller Hinton Agar (MHA) under anaerobic conditions using Gas-Pak (Oxoid) at 37° C. for 72 hrs. Standard inoculums of Optical Density 600 (OD600)=1.0-3.0 were prepared by inoculating colonies of P. acne in Phosphate Buffered Saline (PBS).
- Bacteria Harvesting
-
- 1. The bacterial cells were harvested by adding 1 ml of autoclaved
PBS 1× to the growing bacteria and collected from the plate with a sterile Pasteur pipette; - 2. The bacteria were washed 3 time with PBS;
- 3. The suspension was prepared as 2 McFarland standard/1.5 ml;
- 4. The bacteria were inactivated by heating at 60° C. for 30 min and centrifuged at 5,000×g for 5 min;
- 5. The bacteria were mixed with PBS until reaching the required McFarland/1.5 ml; and
- 1. The bacterial cells were harvested by adding 1 ml of autoclaved
- 1.5 ml of the inactivated bacteria were mixed with 1.5 ml of the Freund's adjuvant complete (SIGMA) and injected subcutaneously into a camel.
- Camel Immunization
- For each infection model, two adult female Arabian camels (Camelus dromedarius) were used; one as control and the other was immunized subcutaneously with an initial dose of 3 ml of prepared Propionibacterium acne vaccine. The vaccinated camel was boosted 4 times at 2 week intervals with 5 ml of the vaccine for each booster. The two camels were kept in a farm for two months, and were kept under similar conditions.
- Detection of Specific Antibodies Against Propionibacterium acne in Milk by Enzyme-Linked Immunosorbent Assay (ELISA)
- For milk antibody screening, the ELISA was performed. Flat bottomed 96 well polystyrene micro titer plates (Greiner, Germany) were coated with 100 μl of 10 μg/m Propionibacterium acne antigens in carbonate-bicarbonate buffer (pH 9.6) overnight at 4° C. The plates were washed 3 times with 100 μl of 0.15 M PBS (pH 7.2) containing 0.05
% Tween 20 and blocked with 100 μl of 2% Bovine Serum Albumin (BSA) in PBS for one hour at Room Temperature (RT). The plates were washed again and 100 μl of the serum samples diluted at 1:100 in 1% BSA were added in duplicates and incubated for one hour at RT, whereas, milk samples were added in duplicates without dilution. Negative and positive control samples were incorporated in each of the plates. After washing, 100 μl of one of the horse radish peroxidase (HRP) conjugated protein A and protein G were diluted at 1:1000 in 1% BSA, and were added separately to each well. The plates were incubated for one hour at RT and washed again. Finally, 100 μl of 0.1% O-phenylenediamine (Sigma, USA) containing hydrogen peroxide in 0.1 M citrate buffer (pH 4.5) were added to each of the wells and absorbance was measured at 490 nm using ELISA reader (AsysHitech, Switzerland). Results are shown inFIG. 1 . - Identification of Camel Milk Proteins Using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoreses (SDS-PAGE)
- Components of camel skimmed milk were fractionated by SDS-PAGE (after precipitation of casein). This was performed using Desaphor VE minigel (Heidelberg, Germany) in the discontinuous buffer system using 0.5 mm thick 10% acrylamide-bisacrylamide gels under non-reducing conditions as described by Hamers-Casterman et al. (1993). For preparing resolving gels, 4 mL distilled water, 2.5 mL running buffer (pH 8.8), 3.3 mL of 30% acrylamide-bisacrylamide solution, 100 μL of 10% ammonium persulfate (APS) and 10 μL Tetramethylethylenediamine (TEMED) were mixed. Stacking gels (4%) were prepared by adding 6 ml of distilled water, 2.5 ml staking buffer (pH 6.6), 1.3 mL of 30% acrylamide-bisacrylamide solution, 100 μl of 10% Ammonium persulfate (APS), and 10 μl TEMED. The milk was mixed with an equal volume of sample buffer lacking B-mercaptoethanol (non-reducing conditions) (pH 6.8). For band size determination, molecular weight protein standard was used after being processed in a similar way as the milk samples. Electrophoresls was carried out using running buffer with pH 8.3 at 120 volts for 60-120 minutes. The gel was stained with Coomassie brilliant blue R-250 (except if used for immunoblot) and destained by 20% acetic acid until clear bands were seen. Results are shown in
FIG. 2 . - In Vitro Activity of Whey Against Acne
- Antibodies isolated from whey of Camel milk immunized with killed P. acne are mixed with a standard inoculum of the bacterium and subsequently incubated for 72 hrs. The total count is determined and then compared with the total count of the standard inoculum without mixing with the antibodies. Results are shown in
FIG. 3 andFIG. 4 . - Preparation of Cream Formula
- For the treatment of acne, a cream formula has been prepared from the following components:
-
Cream Formula 0.5% w/w Material weight mg/unit Camel dried whey* 15 g 2.5 Cetyl alcohol 90 g 15 Tween 2030.9 ml 5.15 Glycerylmonostearate type1 17.1 g 2.85 MERKUR 791 60 g 10 Purified water 387 ml 64.5 Total 600 100 *Equivalent to 0.5% w/w total protein - Preparation Protocol:
-
- 1. Cetyl alcohol, Glyceryl monostearate and Merkur were molten in same container at 75° C. using water bath;
- 2.
Tween 20 was mixed with purified water; - 3. Camel whey powder was dissolved in Tween mixture;
- 4. The mixture from
step 3 was heated to 45°-50° C.; - 5. The mixtures from
step 1 was cooled to 65°-70° C.; and - 6. Both mixtures were mixed together and cooled while mixing until cream turns white.
- Preparation of Gel Formula
- For the treatment of acne, a gel formula has been prepared from the following components:
-
Gel formula 0.5% w/w Material weight mg/unit Camel dried whey* 8.75 g 2.5 Hydroxypropyl cellulose grade HF 10.5 g 3 Deionized (D.I.) water 330.75 ml 94.5 Total 350 g 100 *contains 0.5% total protein. - Preparation Protocol:
-
- 1. Camel dry whey was dissolved in D.I. water;
- 2. Hydroxypropyl cellulose grade HF was dissolved in solution from
step 1; - 3. The mixture was continuously mixed until a clear homogenous gel has formed; and
- 4. The gel was filled in tubes.
- Recovery of Active Ingredients from Cream
- The active ingredients of camel milk whey were re-extracted from the cream for the purpose of stability testing. The cream was mixed with equal volumes of PBS and incubated in water bath at 56° C. for 10 minutes. The mixture was then centrifuged at 13400 rpm for 10 minutes, using microfuge. The aqueous phase was collected and tested using ELISA. Results are shown in
FIG. 5 . - In Vivo Testing of the Efficacy of Whey Protein Concentrate of Milk of Immunized Camel Against Propionibacterium acne
- Protocol:
- Propionibacterium acne (108 CFU/ml (OD600=2)) bacteria were mixed with the whey protein concentrate and incubated for 2 hrs before Intradermal injection in the central portion of rabbit ear. Both positive whey and negative whey were used and compared. Injection was gradually performed by 28 gauge needle to prevent leakage. Gross examination was performed daily along the period of investigation (two weeks). Histopathological evaluation was performed after sacrificing the animals.
- Results:
- Gross Examination:
- Ear thickness: the infection induced rabbit showed double fold increased ear thickness compared to the normal non infected ear. In contrast, ear thickness was significantly reduced in cases of injecting pretreated bacteria with positive whey and less significantly in cases of negative whey. In addition, no significant differences were observed concerning redness, heat, formation of papules and pus.
- Histopathological Examination:
- In the infection induced rabbit, the inflammatory response was presented with dense mixed inflammatory cells and marked eosinophilia, formation of micro abscess and ulcerations without affecting the epidermis. In contrast, mild mixed inflammatory cells without ulceration and less micro abscesses were markedly observed in cases of injecting pretreated bacteria with positive whey. No differences were observed in the negative whey treated rabbit in comparison to the infection induced ear.
- Microbiological Examination
- The recovery of viable P. acne was possible in cases of pus formation. No differences were seen among all groups.
- While the present invention has been described in details and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various additions, omissions and modifications can be made without departing from the spirit and scope thereof.
Claims (16)
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US13/609,342 US10010564B2 (en) | 2012-09-11 | 2012-09-11 | Camel milk-based topical pharmaceutical composition |
US15/986,159 US20180344773A1 (en) | 2012-09-11 | 2018-05-22 | Camel Milk Based Pharmaceutical Composition |
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---|---|---|---|
US13/609,342 Active 2033-05-14 US10010564B2 (en) | 2012-09-11 | 2012-09-11 | Camel milk-based topical pharmaceutical composition |
US15/986,159 Abandoned US20180344773A1 (en) | 2012-09-11 | 2018-05-22 | Camel Milk Based Pharmaceutical Composition |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/609,342 Active 2033-05-14 US10010564B2 (en) | 2012-09-11 | 2012-09-11 | Camel milk-based topical pharmaceutical composition |
Country Status (2)
Country | Link |
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US (2) | US10010564B2 (en) |
GB (1) | GB2507641B (en) |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4324782A (en) * | 1974-01-28 | 1982-04-13 | Beck Lee R | Dental caries inhibiting product of immunized cow's milk having antibodies specific to killed Streptococcus mutans cells |
JP3150162B2 (en) * | 1990-07-05 | 2001-03-26 | 雪印乳業株式会社 | Oral opportunistic infection prevention / treatment composition |
WO1993010818A1 (en) * | 1991-12-04 | 1993-06-10 | New England Medical Center Hospitals, Inc. | Infant formula and infant formula additives |
IL157814A (en) * | 2003-09-08 | 2007-12-03 | Lev Bar Ltd | Cosmetic composition comprising camel milk or components thereof |
WO2008013455A1 (en) * | 2006-07-27 | 2008-01-31 | Lactive B.V. | Means and methods for preventing elevated blood glucose levels in humans with animal antibodies produced in milk |
CN101045750B (en) * | 2007-03-13 | 2010-06-23 | 甘肃省华龙农业开发总公司 | Extraction process of camel colostrum immune globulin IgA, IgG. |
EP2138186A1 (en) * | 2008-06-24 | 2009-12-30 | Nestec S.A. | Probiotics, secretory IgA and inflammation |
GB201003198D0 (en) * | 2010-02-25 | 2010-04-14 | Arab Biotechnology Company | Compositions and methods for the treatment of cancer |
DE102010016134A1 (en) * | 2010-03-25 | 2011-09-29 | Csm Camel Milk Ug | Producing a concentrate from camel milk, useful in pharmaceutical or cosmetic composition for treating skin diseases, comprises preparing cream from camel milk, stirring the cream and separating the camel milk water |
-
2012
- 2012-09-11 US US13/609,342 patent/US10010564B2/en active Active
-
2013
- 2013-09-10 GB GB1316112.0A patent/GB2507641B/en not_active Expired - Fee Related
-
2018
- 2018-05-22 US US15/986,159 patent/US20180344773A1/en not_active Abandoned
Also Published As
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US10010564B2 (en) | 2018-07-03 |
US20140072648A1 (en) | 2014-03-13 |
GB2507641B (en) | 2017-08-30 |
GB2507641A (en) | 2014-05-07 |
GB201316112D0 (en) | 2013-10-23 |
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