TECHNICAL FIELD
The present disclosure relates to a composition comprising stem cell and anti-histamine immunoglobulin and a method to obtain the said composition. More particularly  it relates to method to modulate and or fine-tune the immune system to create the right micro-environment/ niche for stem cell based tissue regeneration  and thus enhancing the success rate of tissue regeneration via stem cell implantations.

BACKGROUND AND PRIOR ART OF THE DISCLOSURE
Anti- histamine immunoglobulins and other similar molecules are used for the treatment of various inflammatory  pain and auto-immune diseases. However  a cure for these conditions still remains an unmet medical need. Applications of stem cells to regenerate the diseased tissue may lead to complete cure. However  the success rates are still poor in this area.

Presently adult mesenchymal stem cells from bone marrow or umbilical cord are being given intra articular in the knee joints for diseases like ankylosing spondylitis  reactive arthritis  filarial arthritis  hypertension  chronic renal failure  chronic articular rheumatism  systemic lupus erythematosus  multiple sclerosis  various types of immunodeficiency syndromes  infection diseases  parasitic diseases  respiratory diseases  autoimmune diseases  eosinophilia caused by malignant tumors  cerbro vascular accident conditions such as Bells palisy  monoplegia  paraplegia  undifferentiated spondylo arthropathies  hypersensitivity  vasculitis  churg-strauss syndrome  rheumatic heart disease  rheumatic heart disease with CCF  rheumatic heart disease with chorea  rheumatoid arthritis  systemic lupus erythematosus  vitiligo  viral encephalitis  drug resistant tuberculosis  acute lepra neuritis  cerebral malaria  primary infertility  meniers diseases  chronic venous ulcers  stress incontinence  chronic cancer pains of hard palet  cervix  chronic cancer pains of secondaries and undifferentiated chronic pains  chronic renal failure  atherosclerosis and chronic pains.
Further  the drugs used are often anti-inflammatory molecules like anti-histamine drugs that do offer a temporary relief in pain or symptoms but does not offer a cure.

The previously carried out studies covered application of anti-histamines in such diseases  whereas  the instant disclosure involves a combination of stem cells and anti-histamine antibodies  showing improved outcome  over the previous studies where anti-histamine was used alone.

The instant disclosure claims for an improved outcome of stem cell based tissue regeneration in the above mentioned diseased conditions  significantly by conditioning the immune system. This manipulation of the immune system is based on our unique observations and understanding of the role of Histamine homeostasis’ of the immune system.

The mechanism of action of the drug histamine conjugated normal immunoglobulin:
Histamine conjugated human immunoglobulin is administered to patients preferably as an injection  subcutaneously or intramuscularly. Intramuscular mode is preferred as its effects are better than subcutaneous injections. When administered intramuscularly  the drug reaches all the lymphnodes and stimulates B cells to produce antihistamine antibodies ("Appa Hista"). This is active process. The antibodies then travel to the inflammatory site and neutralize the histamine  directly or enhance the serum binding ability of the histamine the applicant""s study reveals that the enhancement is nearly 30 times as compared with allergic individuals and helps to reduce the pain  by switching off the (Histamine at site) HI receptors. It may block the specific antibody of IgE synthesis and help the Th1 response to set in the case of intra cellular parasites. It does not interfere with the primary histamine release and does not cause any immunosuppression. Serum binding ability is enhanced by which the threshold and sensitivity to stimuli decreases. The present disclosure attains such high degree of success in the cure of said disease  by administration of a formulation of stem cells and anti-histamine antibiodies or by following a protocol of administering the two entities separately.

Further  it is clear that the release of histamine is a point of the defence strategy is responsible for healing because it gives Th2 type of response. The histamine/1gE/mast cell response is exploited as a strategy by microorganism to cause infections and in turn causes autoimmune diseases. These are also considered as an end result of infection. The inflammation continues as long as histamine is present. The applicant observed that chronic liberation of histamine and Th2 instead of healing damages the healing. The inflammatory site  and the three pathways and their byproducts faci1itate the release of histamine  the histamine positive feedback inturn stimulates IgE Mastcell Pathway and liberates more histamine  and thus it becomes a vicious cycle which is called the "Rao""s Histamine Vicious Cycle". For healing to occur  the cycle must be broken to produce active anti-bodies against histamine so as to enhance the serum binding ability of histamine. The normal individual""s histamine binding ability is thirty times more than that of allergic patients.

The instant disclosure attains such high degree of success in the cure  by administration of a formulation of stem cells and anti-histamine antibiodies or by following a protocol of administering the two entities separately.

STATEMENT OF DISCLOSURE
Accordingly  the present disclosure relates to a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient; a method of obtaining a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipients  said method comprising act of combining the stem cell and the anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipients  to arrive at said composition; a combination comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient  wherein the stem cell and the anti-histamine immunoglobulin are combined in any sequence and after any time interval thereof; a method of preconditioning immune system for improving stem cell therapy in a subject in need thereof  said method comprising act of administering anti-histamine immunoglobulin to the subject  followed by administering the stem cell alternatively with the anti-histamine immunoglobulin for preconditioning the immune system and a method of treating musculo-skeletal conditions  said method comprising step of administering a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient.

BRIEF DESCRIPTION OF THE ACCOMPANYING FIGURES
Figure 1: shows the Rao’s Vicious Cycle
DETAILED DESCRIPTION OF THE DISCLOSURE
The present disclosure relates to a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient.

In an embodiment of the present disclosure  the stem cell is an adult stem cell  selected from group comprising Bone marrow derived mesenchymal stem cell  Bone marrow derived heamatopoetic stem cell  Umbilical cord derived mesenchymal stem cell  Umbilical cord derived haematopoietic stem cell  chondrocyte  osteobalst  neuronal precursor cell  cardiovascular precursor cell and endothelial precursor cell or any combination thereof.

In another embodiment of the present disclosure  the excipient is selected from group comprising granulating agents  binding agents  lubricating agents  disintegrating agents  sweetening agents  glidants  anti-adherents  anti-static agents  surfactants  anti-oxidants  gums  coating agents  coloring agents  flavouring agents  coating agents  plasticizers  preservatives  suspending agents  emulsifying agents  plant cellulosic material and spheronization agents or any combination thereof.

In yet another embodiment of the present disclosure  the composition is formulated into dosage form selected from group comprising tablet  troches  lozenges  aqueous or oily suspensions  ointment  patch  gel  lotion  dentifrice  capsule  emulsion  creams  spray  drops  dispersible powders or granules  emulsion in hard or soft gel capsules  syrups  elixirs  phytoceuticals  nutraceuticals and food stuffs or any combination thereof.

The present disclosure also relates to a method of obtaining a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipients  said method comprising act of combining the stem cell and the anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipients  to arrive at said composition.

The present disclosure also relates to a combination comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient  wherein the stem cell and the anti-histamine immunoglobulin are combined in any sequence and after any time interval thereof.

The present disclosure also relates to a method of preconditioning immune system for improving stem cell therapy in a subject in need thereof  said method comprising act of administering anti-histamine immunoglobulin to the subject  followed by administering the stem cell alternatively with the anti-histamine immunoglobulin for preconditioning the immune system.

In an embodiment of the present disclosure  the anti-histamine immunoglobulin is administered intramuscularly  intra-articularly  intravenous and the stem cell is administered intravenously  intra-artivular  intramuscular  or after loading on biomaterial based scaffolds.

In another embodiment of the present disclosure  the preconditioning is carried out for conditions selected from group comprising bronchitis  rheumatoid arthritis  osteoarthritis  ankylosing spondylitis  asthma  interstitial lung disease  COPD  inflammatory conditions and allogenic stem cell transplantation or any combination thereof.

The present disclosure also relates to a method of treating musculo-skeletal conditions  said method comprising step of administering a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient.

In an embodiment of the present disclosure HISTAGLOBIN is a basic treatment for allergic conditions and not a remedy for acute symptoms. It is therefore recommended for all allergic complaints whatever their symptoms or causes.
(a) Respiratory: bronchial asthma  asthmatic bronchitis  chronic bronchitis  spasmodic tracheitis  hay fever  allergic rhinitis.
(b) Cutaneous: eczema  urticaria  generalised skin allergies.
(c) Nervous: migraine.
(d) Gynaecological: premenstrual syndrome when of allergic aetiology.
(e) HISTAGLOBIN may be used prophylactically with children (where there is a family history of allergy) who are about to undergo an adenoidectomy or tonsillectomy.

In another embodiment of the present disclosure histglobin consists of histamine fixed on an active protein fraction extracted from human blood (gammaglobulin) in strictly defined proportions. It triggers anti-histamine IgG antibodies inside the body and protects the patient from allergy  pain  inflammation etc.

In an embodiment of the present disclosure  the above molecule is used to:
1. Precondition the patients immune system for a better outcome of stem cell therapies
2. Mix this molecule with stem cells in certain propositions and make a new formulation for injection

In an embodiment of the instant disclosure  during inflammation  histamine is released at the inflammatory sites as predicted by Rao’s vicious cycle. The later refers to the proliferation of TH 2 type of reaction and its dominance over TH1 reaction. In this method of cure known as Appa Hista  antihistamine antibodies  combat the histamine at the inflammatory sites and also block TH 2 type of reaction  thus indirectly helping the TH 1 type of reaction. The dual effect of Appa hista antibodies has been visualized in 1) Killing the intracellular organisms through TH 1 type of reaction and; 2) Neutralizing the histamine at inflammation site at the same time blocking the excess TH 2 type of reaction. Since Appa Hista directly attacks histamine where ever it is present  it may cure all inflammatory diseases.

In another embodiment of the present disclosure stem cells are biological cells found in all multicellular organisms  that can divide through mitosis and differentiate into diverse specialized cell types and can self renew to produce more stem cells.

In yet another embodiment of the instant disclosure  in adult organisms  stem cells and progenitor cells act as a repair system for the body  replenished in adult tissues. In a developing embryo  stem cells can differentiate into all the specialized cells  but also maintain the normal turnover of regenerative organs  such as blood  skin  or intestinal tissues.

According to an embodiment of the present disclosure  stem cells can now be artificially grown and transformed into specialized cell types with characteristics consistent with cells of various tissues such as muscles or nerves through cell culture. Highly plastic adult stem cells are routinely used in medical therapies. Stem cells can be taken from a variety of sources  including umbilical cord blood and bone marrow.
In another embodiment of the instant disclosure  bone marrow and umbilical cord mesenchymal and haematopoietic stemcells are now proven for their clinical safety and efficacies (reference). Whartons jelly in the umbilical cord is also a rich source of MSCs. They are also known for their immune modulatory effects and they are usefull for GVHD and for regulating pro-inflamatory cytokines and growth factors (reference). Intravenous injections of these cells in autoimmune diseases like rheumatoid arthritis  injuries  pain  diabetic wounds  parkinsons  cancer etc.  has been reported with low success rates   ie.  in the range of 30 to 40% success per hundred patient studied. In case of arthritis of the knee etc cells have been injected in the joints or given intra-articular.

In an embodiment of the present disclosure  the co-injection of stem cells with histaglobin or administration of the cocktail (of stem cells and histaglobin) given intramuscularly would help in muscle regeneration in Duchen Muscular Dystrophy (DMD) and other musculoskeletal disease patients.

In an embodiment of the current disclosure  the two innovations based on exploitation of the fundamental Rao’s vicious cycle are:
1. “Preconditioning the patients immune system” : A method to modulate and or fine-tune the patients immune system to create the right micro-environment/ niche for stem cell based tissue regeneration  and thus enhancing the success rate of tissue regeneration via stem cell implantations.
2. “ New product formulation” A product involving a combination/ formulation of Adult stem cells (bone marrow or umbilical mesenchymal) and Anti-histamine Immunoglobulins in specific proportions
3.
In an embodiment of the present disclosure the adult stem cell types are selected from a group comprising 1. bone marrow – mesenchymal and heamatopoetic cells. 2. Umbilical cord- mesenchymal and haematopoietic stem cells  3. Differentiated / progenitor cells – like condrocytes  osteobalsts  neuronal precursor cells  cardiovascular and endothelial precursor cells.

The present disclosure is further elaborated with the help of following examples. However  the examples should not be construed to limit the scope of the disclosure.

EXAMPLES

Example 1
Method to precondition the immune system to improve the outcome of stem cell therapies.
The stepwise protocol is provided as below;
Step 1 - Preconditioning the patients immune system:
Dose- 18mg (ie.  one and a half vial)/ each dose (each vial contains 12mg histaglobulin) given once in a week for three weeks   to pre-condition the immune system.

More precisely three injections one each week for 3 weeks   use a cold syringe for every injection  as the product is heat labile
Protocol;
1st injection- histaglobulin- 1st week
2nd injection- histaglobulin- 2nd week
3rd injection- histaglobulin- 3rd week

Step 2 - Injection of stem cells intravenous or intra-articular  or at the site of injuries  by loading on scaffolds  or injection of micro-encapsulated cocooned stem cells for tissue regeneration  wound healing  arthritis and all other indications described in previous patent. Hiatglobulin is injected every 3rd week followed by a stem cell admisntration procedure on the subsequent week.

Protocol;
4th injection –stem cell administration- 4th week
5th injection- histaglobulin- 7th week
6th injection- stem cells administration- 8th week
7th injection- histaglobulin- 11th week
8th injection- stem cell administration- 12th week
9th injection- histaglobulin- 15th week
10th injection- stem cell admisnistartion- 16th week

In an embodiment of the instant disclosure  the preferred dosage of Histamine conjugated gamaglobulin that may be administered intramuscularly may comprise Human Normal Immunoglobulin I.P – 0.25mg  Histamine Dihydrochloride 0.0025 mcg. Or Sodiun Thiosuiphate I.P -0.53 mg.per K.G Body weight. It should be given 3/5 weeks  followed by 3/5 months or once in 21 days by observing various physical and subjective parameters. This is called as one block. During this block inflammatory diseases either may be cured or goes to interval. If the disease reoccurs from the interval the block  it has to be repeated. The route of administration is intramuscuslar.

In an embodiment  the method provided herein is used in allergic and pain conditions like bronchitis  rhematoid arthritis and other related diseases. The patient would be preconditioned by IM (Intramuscular) injections of the Histaglobin  and then  stem cells will be given by IV (Intravenous) route. This is also important and useful in allogenic stem cell transplantation cases  where the foreign/ donor graft will intergate with the host tissue better in a inflammation controlled area.

EXAMPLE 2
Product Formulation
Formulation: Autologus or adult stem cells  or universal donar stem cells like adult pluripotent stem cells in the form of single cell suspension in PBS  suspended in solution in the range of:
- 10 to 100 million cells in 3 ml of PBS can be mixed with 12mg histaglobulin volume 2ml 
- Total volume of preparation- 5ml
- Stored at 4 degree in 5 vials of 1 ml each

Administration: this should be administered IV as short bolus / volumes of 1 ml each at every 10 min.

The total procedure will last over 50 min to 1 hr

In an embodiment  the formulation provided herein is used in musculoskeletal problems generally associated with orthopedics  such as osteoarthritis  ligament tear  fractures  soft tissue injuries and other related diseases.

EXAMPLE 3: Results
Data obtained from Examples 1 and 2 as case studies are compared to data obtained with Control data of injections with Histaglobulins alone  and stem cells alone.

Results obtained on therapies using Method 1 described in Example 1: outcome of preconditioning of immune system with histaglobulins improve stem cell therapy.

Indications studied:
1) Rheumatoid arthritis  2) Osteoarthritis of the knees  3) ankylosing spondylitis
Therapy: Method: same as described in example 1 above for histaglobulin and stem cell injections. Autologus or allogenic mesenchymal stem cells were given intravenously (IV) at 2million cells/ kg body weight in all the diseases/ medical conditions.

Results: Table 1
Indications No of Patients Histaglobulin only Stem cells only Histaglobulin plus stem cells
Rheumatoid arthritis Enrolled
25 20 24
Recovery
15 (60%) 13 (65%) 21 (87.5%)
Osteoarthritis of the knees Enrolled
24 16 10
Recovery
15 (62%) 10 (62.5%) 8 (80%)

Discussion
Histaglobin plus stem cells injected following the instant protocol gives significantly better recovery.
More disease conditions and patients need to be studied

Results obtained on therapies using the formulation/ product [Method 2] described in Example 2.

Indications studied: 1) Rheumatoid arthritis  2) osteoarthritis 3) ILD etc
Therapy: as described in method 2 above

Results: Table 2
Indications No of Patients Histaglobulin only Stem cells only Histaglobulin plus stem cells
Rheumatoid arthritis Enrolled
15 9 5
Recovery
10 (66%) 5 (55%) 4 (80%)

Discussion
Injection of the new formulation gives improved results  and the outcome is comparable to Method 1 in efficacies.

We Claim:

1. A composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient.
2. The composition as claimed in claim 1  wherein the stem cell is an adult stem cell  selected from group comprising Bone marrow derived mesenchymal stem cell  Bone marrow derived heamatopoetic stem cell  Umbilical cord derived mesenchymal stem cell  Umbilical cord derived haematopoietic stem cell  chondrocyte  osteobalst  neuronal precursor cell  cardiovascular precursor cell and endothelial precursor cell or any combination thereof.
3. The composition as claimed in claim 1  wherein the excipient is selected from group comprising granulating agents  binding agents  lubricating agents  disintegrating agents  sweetening agents  glidants  anti-adherents  anti-static agents  surfactants  anti-oxidants  gums  coating agents  coloring agents  flavouring agents  coating agents  plasticizers  preservatives  suspending agents  emulsifying agents  plant cellulosic material and spheronization agents or any combination thereof.
4. The composition as claimed in claim 1  wherein the composition is formulated into dosage form selected from group comprising tablet  troches  lozenges  aqueous or oily suspensions  ointment  patch  gel  lotion  dentifrice  capsule  emulsion  creams  spray  drops  dispersible powders or granules  emulsion in hard or soft gel capsules  syrups  elixirs  phytoceuticals  nutraceuticals and food stuffs or any combination thereof.
5. A method of obtaining a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipients  said method comprising act of combining the stem cell and the anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipients  to arrive at said composition.
6. A combination comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient  wherein the stem cell and the anti-histamine immunoglobulin are combined in any sequence and after any time interval thereof.
7. A method of preconditioning immune system for improving stem cell therapy in a subject in need thereof  said method comprising act of administering anti-histamine immunoglobulin to the subject  followed by administering the stem cell alternatively with the anti-histamine immunoglobulin for preconditioning the immune system.
8. The method as claimed in claim 7  wherein the anti-histamine immunoglobulin is administered intramuscularly  intra-articularly  intravenous and the stem cell is administered intravenously  intra-artivular  intramuscular  or after loading on biomaterial based scaffolds.
9. The method as claimed in claim 7  wherein the preconditioning is carried out for conditions selected from group comprising bronchitis  rheumatoid arthritis  osteoarthritis  ankylosing spondylitis  asthma  interstitial lung disease  COPD  inflammatory conditions and allogenic stem cell transplantation or any combination thereof.
10. A method of treating musculo-skeletal conditions  said method comprising step of administering a composition comprising stem cell and anti-histamine immunoglobulin  optionally along with pharmaceutically acceptable excipient.