Oxygen carriers for the treatment of skin indispositions

Abstract

The present invention refers to an oxygen carrier, preferably hemoglobin or myoglobin, for use in the treatment of an unpleasant feeling of a body surface due to a disease or a disorder.

Claims

1. A method for treating pain, itching, stinging, a burning feeling, numbness or swelling of skin of a patient due to a disorder or disease affecting the somatosensory nervous system, the method comprising applying a composition to skin of the patient, the composition comprising (a) an oxygen carrier having at least 40% of oxygen-binding sites charged with a non-O.sub.2 ligand, and (b) at least one further ingredient, selected from electrolyte(s) preservative(s), stabilizer(s), anti-flocculant(s), anticoagulant(s), pH buffering agent(s), solvent(s), antioxidant(s), film-forming agent(s), ointment ingredients, oils, greases or fats, or crosslinking agent(s).

2. The method according to claim 1, wherein the composition is an aqueous or organic solution, a lotion, a cream, a gel, an ointment or a powder, or said composition is comprised in a plaster, bandage or dressing.

3. The method according to claim 1, wherein the composition comprises: (i) between about 40% to about 90% of the oxygen carrier provided in a form wherein the oxygen-binding sites are charged with a non-O.sub.2 ligand, (ii) the oxygen carrier is a naturally occurring hemoglobin or myoglobin of human or animal origin, or is artificially treated, crosslinked or modified hemoglobin or myoglobin of human or animal origin; and (iii) the composition is provided in sterilized form.

4. The method according to claim 1, wherein the pain, itching, stinging, burning, numbness or swelling of a patient due to a disorder or disease occurs at body sites or body areas showing blebs, blisters, pimples, rash, red patches, or wheals.

5. The method according to claim 1, wherein the composition is provided in a packaging means comprising a container, a tube, a can, an aerosol can, airless dispenser, airless pump dispenser, airless tube, a spraying container or spraying flask, as far as such devices work without sucking back air after an application use, a one-unit-package, an ampulla, a pouch, sachet, package or blow-formed container.

6. A method for preparing a composition for use according to claim 1, comprising: (i) charging the oxygen carrier with the non-O.sub.2 ligand during or after isolation from its natural environment, (ii) adding at least one further ingredient (b); (iii) sterilizing the composition by one or more of heating, filtration, centrifugation, addition of preservatives, vapour application, gas application or UV-application or any combination thereof; and (iv) packaging the composition.

7. The method of claim 6, wherein the oxygen carrier compound is isolated and purified from whole blood from a human, animal, or mammal.

8. The method according to claim 1, wherein the oxygen carrier is a naturally occurring hemoglobin or myoglobin of human or animal origin.

9. The method according to claim 1, wherein the non-O.sub.2 ligand is selected from carbon monoxide or nitrogen monoxide.

10. The method according to claim 1, wherein the disorder or disease is caused by a virus.

11. The method according to claim 10, wherein the disorder or disease is caused by a Herpes virus, a Herpes Zoster virus, a Herpes simplex virus, shingles, chickenpox, or lip blisters.

12. The method according to claim 1, wherein the composition is provided in the form of a lotion, cream, or ointment and further comprises a cosmetically-acceptable oil, wax, grease, or fat commonly used in cremes, lotions, emulsions or ointments.

Description

EXAMPLES

Example 1: Preparation of the Oxygen Carrier and a Composition Comprising Same

(1) Hemoglobin was isolated from whole blood of pigs by separating the red blood cells from serum, lysing the collected red blood cells, pelleting cell debris, charging the hemoglobin with CO by introducing CO gas until saturation of the liquid sample is obtained, heating the solution, carrying out several filtration steps, including a virus filtration step and washing the obtained hemoglobin solution by adding twice a 2-fold volume of 0.9% saline and filtering the solution.

(2) A ready-to-use composition for treatment of a body surface was prepared, comprising 10% of purified and stabilized hemoglobin, 0.05% N-acetyl cysteine and 0.7% phenoxy ethanol in 0.9% NaCl. The composition was charged again with CO gas, separated into 10 portions and packaged into an aerosol can, respectively.

Example 2: Comparison of Charged/Non-Charged Hemoglobin

(3) A) A charged hemoglobin-spray was prepared according to Example 1

(4) A ready-to-use composition for treatment of a body surface was prepared, comprising 10% of purified and stabilized hemoglobin, 0.05% N-acetyl cysteine and 0.7% phenoxy ethanol in 0.9% NaCl. The composition was charged again with CO gas, separated into 20 portions and packaged into an aerosol can, respectively.

(5) The composition can be stored between 4° C. and room temperature for months to years.

(6) B) A second portion of the pig whole blood was treated as in Example 1 with the exception that no CO charging was carried out. During the heating step a considerable amount of hemoglobin precipitated. The following filtration steps were difficult to carry out. The yield of purified hemoglobin per liter whole blood decreased to less than 30% of the yield when the CO charging is carried out before heating. This shows that charging the samples with CO during preparation stabilizes the hemoglobin in a way that during a heating step considerably less hemoglobin precipitates.

(7) A ready-to-use composition for treatment of a body surface was prepared, comprising 10% of purified hemoglobin, 0.05% N-acetyl cysteine and 0.7% phenoxy ethanol in 0.9% NaCl. The composition was portioned and packaged into aerosol cans, respectively.

Example 3

(8) Samples of compositions prepared according to Example 2A) or Example 2B), respectively, were examined considering the O.sub.2 charge of the samples as well as the production of methemoglobin, a species which is unable to bind O.sub.2. Furthermore methemoglobin affects hemoglobin molecules in the immediate vicinity in a way that these can still bind O.sub.2, but cannot release it no more.

(9) All the samples of Table 1 were diluted 1:1 with a 0.9% NaCl solution. Samples 3.1 to 3.5 were measured immediately after dilution (storing of samples 3.4 and 3.5 was at room temperature before dilution). Gas treatment was as follows: 6 ml of said diluted solution were transferred into a 30 ml glass flask each. Gas treatment was carried out by filling the glass flask completely with the respective gas, closing the flask and pivoting the sample for 30 sec. All the samples were stored for the mentioned time period at 30° C. The samples were treated according to the conditions described in Table 1 and the total amount of hemoglobin (hemoglobin+methemoglobin), the oxygen content, the methemoglobin content and optionally the CO content was measured.

(10) TABLE-US-00001 TABLE 1 totalHb Example sample treatment g/dL O.sub.2 % CO % MetHb % 3.1 Freshly prepared comp. none 5.5 1.7 96.3 3.9 according to Example 2A) 3.2 Freshly prepared comp. none 5.7 26.8 5.1 According to Example 2B) 3.3 Composition of Example 2A), none 5.9 1.8 96.5 3.8 stored for 3 month (10° C.) 3.4 Composition of Example 3.2 Stored for 24 h 5.6 26.9 11.2 3.5 Composition of Example 3.2 Stored for 48 h 5.5 27.0 17.0 3.6 Composition of Example 3.3 0.5 h O.sub.2 5.8 13.7 84.7 4.4 3.7 Composition of Example 3.3 1 h O.sub.2 5.7 19.6 79.1 4.2 3.8 Composition of Example 3.3 2 h O.sub.2 5.8 25.7 73.1 4.3 3.9 Composition of Example 3.3 3 h O.sub.2 5.9 30.7 68.1 4.2 3.10 Composition of Example 3.3 24 h O.sub.2 5.7 33.5 59.6 9.8 3.11 Composition of Example 3.3 48 h O.sub.2 5.8 32.5 55.9 15.5 3.12 Composition of Example 3.3 72 h O.sub.2 5.7 31.7 55.8 16.0 3.13 Composition of Example 3.3 0.5 h CO.sub.2 5.8 4.0 91.8 5.7 3.14 Composition of Example 3.3 1 h CO.sub.2 5.7 6.0 90.0 5.7 3.15 Composition of Example 3.3 2 h CO.sub.2 5.9 9.2 86.7 5.8 3.16 Composition of Example 3.3 3 h CO.sub.2 5.7 9.6 85.1 6.9 3.17 Composition of Example 3.3 24 h CO.sub.2 5.6 4.9 77.5 19.7 3.18 Composition of Example 3.3 48 h CO.sub.2 5.8 8.2 68.2 27.4 3.19 Composition of Example 3.3 72 h CO.sub.2 5.7 9.4 65.9 28.3 3.20 Composition of Example 3.3 0.5 h air 5.8 9.2 89.0 4.5 3.21 Composition of Example 3.3 1 h air 5.7 12.4 85.9 4.4 3.22 Composition of Example 3.3 2 h air 5.9 18.0 80.5 4.3 3.23 Composition of Example 3.3 3 h air 5.6 20.7 77.9 4.3 3.24 Composition of Example 3.3 24 h air 5.8 24.3 69.3 9.1 3.25 Composition of Example 3.3 48 h air 5.7 25.4 64.3 13.9 3.26 Composition of Example 3.3 72 h air 5.7 26.8 62.2 14.3

(11) As can be seen from the results in Table 1 the compositions of the invention, wherein the oxygen carrier is charged with CO, not only can be stored for a long time without forming methemoglobin, but further are able to replace the bound CO by O.sub.2 when it is offered to the charged oxygen carrier. If exposed to 100% O.sub.2 (examples 3.6 to 3.12) the O.sub.2 saturation of the hemoglobin increases very fast.

(12) If the composition is exposed to CO.sub.2, representing the situation inside of mammalian pathway-active tissue, an increased amount of methemoglobin is formed (examples 3.13 to 3.19).

(13) As can be seen the content of O.sub.2 in freshly prepared samples (Example 3.2) is around 27%. Said content remains stable during storage (see Examples 3.4 and 3.5), however, the methemoglobin content increases undesirably. Considering Examples 3.20 to 3.26 the surprising result is that within 3 hours the CO comprising composition exposed to air is charged with 20.7% oxygen, but still has a very low methemoglobin content. When a body surface is treated, the composition is applied to the (cleaned) surface and remains in contact with air.

(14) These results show that a composition according to the invention comprises a stabilized oxygen carrier which after several months of storing provides high oxygen transport when it is in contact with air.

Example 4

(15) In total 100 persons suffering from a Herpes zoster infection (Shingles) and reporting about painful feelings where divided into two groups of 50 patients each, which in view of severity of the disease, age and sex of the patients and further conditions were comparable.

(16) All the patients of both groups were treated by obtaining a classic standard Shingles treatment therapy (oral application of an antiviral medicament). In Group 1 the patients additionally were treated by spraying a sterile saline solution as a thin film to the body areas affected by Shingles. In Group 2, however, the patients were treated by spraying a thin film of a composition as prepared according to Example 1 to the body areas affected by Shingle blisters (“Standard of Care” plus Granulox® application at each treatment of shingles). Spray applications were carried out twice a week by topical application of the hemoglobin spray Granulox® (Sastomed, Georgsmarienhütte, Germany), a composition according to Example 1.

(17) All the patients were treated until the visible syndromes were healed and the unpleasant feeling (itching, stinging, pain etc.) was overcome.

(18) Data were collected twice weekly for four weeks then weekly until the patient was discharged (healed/Pain free) or at 12 weeks stage where the study data collection and patient follow up will cease.

(19) Pain reduction over time (Visual analogue score 0=no pain, 10 =maximal pain) and time to heal was evaluated by questionnaires with scoring tools.

(20) Group 1: “Saline group”: 50 patients

(21) Male: 29

(22) Female: 21

(23) Age range: 16 to 89 years

(24) Pyrexia on day 0: 23 patients

(25) Rash: 9

(26) Blister: 35

(27) Erythema: 3

(28) Other: 3 (pain only)

(29) Oral Aciclovir medicaments as standard: 49

(30) (1 patient had allergy)

(31) Self care with skin wash saline: 50

(32) Pain range in days: 15 to 59

(33) Healing days: 5 to 43

(34) Group 2: “Granulox® group”: 50 patients

(35) Male: 26

(36) Female 24

(37) Age range: 17 to 90 years

(38) Pyrexia on day 0: 26 patients

(39) Rash: 3

(40) Blister: 36

(41) Erythema: 5

(42) Other: 6 (pain only)

(43) Oral acyclovir application: 50

(44) Self care with Granulox®: 50

(45) Pain range in days: 9 to 30

(46) Healing days: 5 to 18

(47) As can be seen in the above results the time period of painful feelings elicited by the underlying disease noticeably decreases when the patients were treated with the hemoglobin spray Granulox® instead of saline. Also the visible symptoms (blisters, red dots) are much faster reduced when Granulox® was applied to the affected body areas. Thus, due to the positive effect of the hemoglobin spray the healing time was impressively shorter.