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Spleen Cytokines Against Inflammatory Bowel Disease

Vladimir N Pak, Vladimir Pak received his Ph.D. from the Institute of Bioorganic Chemistry, Moscow. He modernized technologies and manufactured APIs for known drugs. Invented anticancer drug candidates based on alpha-fetoprotein that can be used as injectable or orally. Developed supplements to treat stomach ulcers and IBD.

Spleen cytokines regulate the immune responses through multiple pathways, contributing to a normal function of the immune system. They have been successfully used to treat and heal stomach ulcers in both humans and horses. As a tablet, nutritional supplement, and/or an enema, spleen cytokines can be used against diseases with immunologic insufficiency or imbalance, including inflammatory bowel disease.

Over one hundred years ago, at the University of Toronto, Frederick Banting, Charles Best, and James Collip extracted protein insulin from the pancreas that saved the lives of millions of diabetics since then. It is possible, that the proteins extracted from the spleen can also help people with inflammatory bowel disease (IBD).

The spleen is an organ found in all vertebrates. It is a large secondary lymphoid organ that primarily functions as a blood filter. The second spleen’s function is immune system surveillance. As a specialized immune organ, the spleen plays a significant role in innate and adaptive immunity. Specifically, the spleen is a crucial organ for peripheral immune tolerance, complementing central immune tolerance. Splenectomy in both study animals and humans results in overwhelming infections with especially, encapsulated bacteria [1]. On the opposite, supporting the immune system with an additional spleen or the substances it produces can help to treat many diseases. In world practice, there are some commercial preparations of the spleen extracts, such as Solkosplen, Splenic, Polygram, Splenopid, Splenaktiv, and others [2] that have been liscensed for use to treat diseases.

One spleen is good and two are better

The spleen’s ability to neutralize pathogens (especially encapsulated bacteria) through phagocytosis was first tested under clinical conditions. The porcine spleen was chosen as the most immunologically close to the human one. The fresh isolated porcine spleen was connected to the patient’s blood circulation. Perfusion of the patient’s blood through the spleen was effective to treat sepsis, rheumatoid arthritis, lupus erythematosus, and other diseases [3-5]. This perfusion method proved to be complicated [6] and spleen extracts were developed and used as treatment alternatives. The biologically active substances produced by the spleen affect different parts of the immune homeostasis. Spleen cells produce opsonins (tuftsin, fibronectin, etc.), and the whole complex of cytokines that coordinate the activity of innate and adaptive immune cells. Cytokines are small proteins that affect the growth of all blood cells and other cells that help the body's immune and inflammation responses.

Spleen protein fractions with molecular weight <74 kDa have demonstrated antibacterial and immune-stimulating activity. Splenopid and Splenaktiv injectable drugs were used to coordinate the immune response during diseases with immunological insufficiency: ulcers, sepsis, autoimmune diseases, oncological pathologies, diabetic foot, complex treatment of multiple sclerosis, etc. [2, 7-9].

The level of pro-inflammatory, anti-inflammatory cytokines, and interferons in Splenaktiv exceed the serum levels in the blood of healthy donors. Apparently, Splenaktiv can cause a change in the state of immune effectors. Because of the stimulation of hematopoiesis their amount increases, the amount of differentiated subpopulations increases, and its functional activity stimulates. Primarily, these processes occur in the immunoregulatory system of T-lymphocytes, preferentially in a subpopulation of Th1. Splenaktiv as the first phase cytokine complex of the immune response can modulate the activity of immune effectors of this phase of macrophages, Th1-lymphocytes, and neutrophilic granulocytes [10].

The reverse-phase HPLC of Splenopid shows two majors proteins peaks (Fig. 1).

Figure 1. The HPLC of Splenopid (A) and Omega Alpha Pharmaceuticals Inc. spleen concentrate (B). The two major marker proteins are in the red circles.

Some of the Splenaktiv and Splenopid proteins have been identified using electrophoresis and ELISA to be: IL-1, IL-2, IL-3, IL-4, IL-10, receptor IL-1b antagonist (IL1-RA), TNF-α, IFN-γ, GM-CSF, and others. Out of 176 mg of Splenaktiv (or 230 mg of Splenopid) in a vial, interleukins represent only mcg quantities [10, 11].

In a study of patients with stomach ulcers, Splenopid was injected around the stomach ulcers using an endoscope. The treatments were very effective and 148 patients remained ulcer free for a long time [5, 12]. Nevertheless, this technique is highly invasive and complicated, and it is an expensive personalized medicine.

When you ask what patients want - obviously, they want appropriate therapy that works. But if you ask if they want an infusion or an oral drug, most would want an oral drug.

In another study the tumor-bearing mice were gavaged with the porcine spleen extract. The extract inhibited tumor growth and enlarged survival rate in mice to 70% compared to 0-10% in control groups [13, 14]. Inhibition of tumor growth proves that the peroral spleen extract can be used orally instead of in injections. The spleen proteins are a safe natural product to be used by the oral route, and they can be helpful in treating patients with different pathologic processes connected to the suppression of cellular and humoral immunity which needs immune correction. For example, the supplement with a complex set of oligo- and polypeptides constituted of 163 different spleen peptides with molecular weight <10 kDa can increase Th1 responses and decrease Th2 responses, providing an adequate balance for the immune responses [15].

On the other hand, if the mucous membranes are damaged, spleen proteins can work locally interacting directly with the exposed and affordable lymphoid and blood cells.

Omega Alpha Pharmaceuticals Inc. has developed technology for manufacturing the spleen cytokines concentrate which has the same two major protein markers as in Splenopid (Fig. 1) and has tested it in horses.

According to a study done at the Western College of Veterinary Medicine in Saskatchewan, 74.5% of the 94 racehorses involved in the study had stomach ulcers because of the permanent stress condition during races [16]. The existing drugs, for example, omeprazole, do not heal ulcers, and clinical signs return in all cases. Omega Alpha Pharmaceuticals has tested in 2 separate trials the effects of orally administered spleen extracts on stomach ulcers seen in equine athletes.
Cytokines act better and faster than existing drugs, heal and prolong relief of stomach ulcer disease, irritable bowels, and irritable bowel syndrome, and provided a stable effect. The beneficial effect with a long remission period was confirmed by endoscopic examination (Fig. 2).

Figure 2. Ulcer wounds with red blood spots are present Before and absent After the treatment with spleen cytokines.

There were some promising results in horses that suggest that spleen cytokines may have also a beneficial effect on colonic ulcers. Some anecdotal evidence obtained with ultrasound suggests that they would be effective in helping to alleviate the symptoms associated with IBD.

IBD is a broad term that describes conditions characterized by chronic inflammation of the GI tract. This chronic and prolonged inflammation impairs the ability of affected organs to function properly, leading to symptoms such as persistent diarrhea, abdominal pain, rectal bleeding, weight loss, and fatigue. Crohn’s disease and ulcerative colitis are the two most common IBDs. Crohn’s disease can affect any part of the GI tract, from the mouth to the anus. It is a lifelong condition that can impact a person’s quality of life significantly. Ulcerative colitis is limited to the large intestine (colon) and the rectum. The inflammation occurs only in the innermost layer of the lining of the intestine. In people with IBD, the immune system mounts an inappropriate response to the GI tract, resulting in inflammation.

About 300,000 Canadians are living with IBD. There is no cure for IBD, and the treatment usually involves either anti-inflammatories, immunosuppressants, biologics, and antibiotics therapy or surgery. The goal of treatment is to reduce the inflammation that triggers the signs and symptoms, thus providing relief as well as long-term remission and reduced risks of complications. The current treatment paradigm for IBD is to use biologic therapy early to achieve clinical remission and mucosal healing, which will ultimately decrease the risk of corticosteroid use, surgeries, and hospitalizations and increase the quality of life.

The cost of IBD care is rising worldwide as IBD incidence and prevalence are rapidly increasing. Anti-inflammatory molecules, particularly biologics (anti-TNF, Stelara, etc.) are expensive.

The new suppression mechanism of inflammation through modulation of monocyte/macrophage activity was demonstrated with alpha-fetoprotein (AFP). Previously, the natural AFP was successfully injected to treat patients with IBD [17]. In an animal model recombinant AFP has shown the results similar to anti-TNF-α which is a “Gold Standard” in the IBD treatment [18].

One spleen is good and seven are better.

While monocytes/macrophages are at the top of the immune cell hierarchy and can be affected by nutrients specifically delivered by AFP [19, 20], cytokines are mediators in the whole immune cells network. It is known that diseases with the development of secondary immunodeficiency cellular link and macrophage system, lead to the imbalance of cytokines, which have to be corrected by cytokine administration. The cytokines administration can shift the balance to a normal/calming ratio leading to success during the treatment of a lot of diseases that need immune correction.

The Omega Alpha Pharmaceuticals Inc. supplement contains the reduced “horse” dose of the spleen cytokines (7 times concentrate) confirmed by the two HPLC markers of Splenopid. It can be manufactured in normal or in acid-resistant capsules/tablets. Normal capsules dissolve in acid conditions and can be used for stomach ulcers and Crohn’s disease treatment. An acid-resistant capsule can move through the stomach and calm the large intestine inflammations. In several anecdotal cases, cytokines in capsules were used to treat stomach ulcers in patients. They have demonstrated simultaneously antibacterial and healing activity similar to Splenopid injections (unpublished).

On the other hand, an enema (a clyster) is an easy way to access an inflamed colon. Though clyster is an unpleasant procedure, surgery is much more disturbing. Previously, the best results in colitis treatment in 116 patients were achieved with combined traditional treatment (salazopiridazine, glucocorticoids, immunosuppressants, and antibiotics) and rectal infusions of Splenopid by a clyster. Splenopid infusions had a beneficial effect on colonic ulcers: the disappearance of ulcer bleeding in 39% of patients. Microscopic examination of the biopsy material of the colon mucosa in all patients receiving complex treatment revealed a decrease in the inflammatory process, expressed in a decrease in the inflammatory infiltrate, in the disappearance of crypt abscesses, epithelialization of ulcerative defects. Ulcerative defects decreased in size with the initial signs of epithelialization, and the crypts of the mucosa shortened and deformed. The was carried out from four months to three years. Only a slight decrease in inflammatory infiltrate was registered in 30 patients of the control group. The complex method of treatment of patients with chronic nonspecific ulcerative colitis using Splenopid made it possible to optimize the tactics of treatment of patients with the widespread nature of lesions of the colon mucosa [21, 22].

Unlike Splenopid or other injectables, cytokines for capsules or a clyster do not need high purity and sterility making the manufacturing cheap. Also, they do not provoke adverse reactions in high doses. The concentrated cytokines complex has a more powerful action than mono biologics. The spleen’s cytokines concentrate can be both as prevention of IBD flare-ups and also as a much more cost-effective alternative treatment to the currently available biologics. Clinical trials are needed to test the spleen’s cytokines concentrate doses/courses in IBD treatment in an enema, capsules, tablets, suppository, and other forms.

Conclusion

Drugs with spleen cytokines can coordinate the immune response during diseases with immunological insufficiency or misbalance: ulcers, sepsis, autoimmune diseases, IBD, oncological pathologies, etc. The spleen cytokines concentrate is expected to be effective in IBD treatment in an enema, capsules, tablets, suppository, and/or other forms.

P.S. Botanicals and L-glutamine, which also reduce inflammation and support the healing of the gut were added to the spleen cytokines concentrate. With the NPN 80070540 supplement Litis is registered by Health Canada.

References:

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Vladimir N Pak

Research Scientist, Omega Alpha Pharmaceuticals Inc

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