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NOVEL PEPTIDE
by Dr. James Bond
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Introduction
Currently, many biological response modifiers (BRM) have been identified. Examples are interleukins
and cytokines. Also, bone marrow plays an important role in the overall immunomodulation. One
could say that bone marrow is the brain of the immune system.
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In 1560 Tisulis made a first description of bone marrow, but it took almost four centuries until in
1936, Bummar suspected that bone marrow plays an important role in the immune system after birth.
Today, bone marrow is considered to have a key function in the development and function of the
immune system and the biological defense mechanisms against cancer and chronically infected cells.
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Bone marrow and its effects on the immune system and lymphatic tissue
Bone marrow tissue is responsible for selected transformation of precursor cells into different B-cells,
i.e. helper B-lymphocytes, which aid in the differentiation of other lymphocytes, killer cells (NK
cells), cytotoxic cells, and suppressor B-lymphocytes (1-3). Having been released into the blood
stream, intestinal and peripheral tissues, the B-lymphocytes are characterized by well-defined antigens
or activation markers on their surface.
The bone marrow is directly innervated, thus making its role in the interaction between the immune
system and the neuro-endocrinal systems understandable. In newborn mice, removal of bone marrow
causes a significant change and decrease in lymphatic tissue and a hypofunction of the ReticuloEndothelial System (RES). In addition, the maturation of B-dependent lymphocytes is severely
hampered, or even made impossible.
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Bone marrow and its effects on haematopoietic factors
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There is a delicate interaction between inactive bone marrow and active bone marrow. There is a direct
and positive correlation between hypofunction of bone marrow and the decline of production of
colony stimulating factors (CSF). Therefore, in cases where there is insufficient production of CSF,
the therapeutic application of bone marrow peptides can be helpful.
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The Invention
BREAKTHROUGH!
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We have isolated a tissue specific Immuno regulator peptide from human bone marrow, named
"B007".
We screened, using PCR, 16 human cDNA libraries from 16 different tissues and found B007 cDNA
only in human bone marrow.
We screened the Human, Mouse and Rat ESTs bank and found no similarity to any known gene. We
got similar results for proteins and total genomic DNA banks, so B007 is not known.
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Using conventional bioinformatic tools, we found that B007 has the following characteristics:
(1) it is soluble,
(2) it has a signal peptide,
(3) it has a total secondary structure like that of Growth Hormone (GH).
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The combination of tissue specificity and physical characteristics of a peptide hormone renders B007 a
very attractive candidate for use in promoting the cellular immune response and, accordingly, for
developing a drug to treat different clinical conditions such as HIV, Chronic hepatitis B (CHB),
Chronic hepatitis C (CHC), Malaria, etc (see below).
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The B007 peptide encoded by the cDNA is 84 amino acids long and includes a signal peptide of 33
amino acids on its N-terminal end. The cDNA sequence and amino acid sequence of B007 are as follows:
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atgctggcattcagaagccaggggctcatgttaccccaga
gctgcccacaactggctatcctcaccctaagtgccttggc
agcagtgtcttgttcagctctgcatctctggcttagtggg
gagccagtccagagctctgcaacaaaggacatgagatcca
aatccgattccaagcgagtgagtgacaagcagctaatttc
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caaagctgtgtggtggacattttttcttccttcaaccctc
tgggagagaaaatga
MMALLSQKLMLPQSCPQLKFLTLSALAAVSFSALHEWLSG
EPVQSSGTKDMRSKSDSKRVSDKQLISKAVWWTFFLPSTE
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WERK
The B007 peptide includes a 33-amino acid signal sequence. Thus, the peptide without the
signal peptide consists of the following sequence:
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LHEWLSGEPVQSSGTKDMRSKSDSKRVSDKQLISKAVWWT
FFLPSTEWERK
The nucleic acid molecule encoding the above amino acid sequence has the following
sequence:
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catctctggcttagtggggagccagtccagagctctggaacaaaggacatgagatc
caaatccgattccaagcgagtgagtgacaagcagctaatttccaaagctgtgtggt
ggacattttttcttccttcaaccctctgggagagaaaatga
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The following partial B007 peptides were found to also have biological activity (see
experimental results reported below):
a) SGEPVQSSGTKDMRSKSDSKRVS
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b) DKQLISKAVWWTFFLPSTEWERK
c) PSTEWERK
d) AVWWTFFLPSTEW
e) AVWWTFFLPSTEWERK
f) WTFFL
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Sequence (a) consists of amino acids 6 to 28 of the B007 peptide; Sequence (b) consists of
amino acids 29 to 51 of the B007 peptide; Sequence (c) consists of amino acids 44 to 51 of the
B007 peptide; Sequence (d) consists of amino acids 36 to 48 of the B007 peptide; Sequence (e)
consists of amino acids 36 to 51 of the B007 peptide in the reverse order; and Sequence (f) consists
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of amino acids 39 to 43 of the B007 peptide.
Indications also exist that L-amino acids may be replaced by D-amino acids without
affecting activity, or perhaps even increasing the activity.
Experimental Results
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Several deletion mutants of the B007 peptide were constructed and synthesized. These
mutants were tested for their ability to stimulate proliferation in human peripheral blood
lymphocytes (hPBL) as compared with the wild-type B007 peptide (WT). The right-hand column
indicates the fold increase in proliferation over proliferation in untreated cells.
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TABLE 1: Stimulation of Proliferation in Human PBL
Peptide
Amino acid sequence
Fold
Increase
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Sequence (a)
LHEWLSGEPVQSSGTKDMRSKSDSKRV
SDKQLISKAVWWTFFLPSTEWERK
SGEPVQSSGTKDMRSKSDSKRVS
Sequence (b)
DKQLISKAVWWTFFLPSTEWERK
2.2-2.8
Sequence (c)
PSTEWERK
1.1-1.3
Sequence (d)
AVWWTFFLPSTEW
2-2.2
Sequence (e)
AVWWTFFLPSTEWERK
2-2.4
WT
1.4-1.6
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Clinical applications of bone marrow peptides
The opportunity
Most clinical conditions arising from chronic infections of viruses, fungi, yeast and parasitic infections
have only very limited remedies. HIV, CHB, CHC and malaria are chronic diseases from which tens
of million of people suffer nowadays without any cure.
The immune system is responsible for vigilance against these chronic virus, fungi, yeast, and parasitic
infections, as well as neoplasms and aging. Bone marrow extracts have been used clinically in a
variety of ways involving some of these conditions. They have been used orally and as injectables; by
themselves and in combination with other therapeutics. Bone marrow extracts have been used to treat
severe and chronic allergies involving the respiratory tract and skin as well as in severe acute and
chronic infectious diseases. The extracts have also been shown to reduce post surgical infections,
decrease the damage of chemotherapy and radiation and have been used as adjuncts to mainstream
therapy for treatment of neoplasms.
All of these conditions were treated successfully with bovine bone marrow extracts. The market for
treatment of these conditions is huge and waiting for something more specific and powerful than
extracts from bovine bone marrow.
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Proposed Examples for Future Clinical trials with Bone marrow peptides
 Administrating bone marrow peptides to patients with malignant melanoma, and following
tumor-free period, survival time and quality of life.
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 Administrating bone marrow peptides to patients with intermediate- and high-grade NonHodgkins lymphoma, in addition to standard chemotherapy, and following toleration of the
bone marrow peptides by the patients, and response rate as compared to patients who did not
receive bone marrow peptides.
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 Administrating bone marrow peptides in addition to the drug cefotetan to patients undergoing
colorectal surgery and following the rate of abdominal abscesses and upper respiratory tract
infections.
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 Administrating bone marrow peptides to women with advanced breast cancer in addition to their
chemotherapy regimen, and following toleration of the bone marrow peptides by the patients as
well as the rate of secondary infections.
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It is believed that the bone marrow peptides described above (and possibly many other variants of the
B007 peptide) may be used:
1. to enhance bone marrow function and protect the patient against myelo-suppression of standard
chemotherapy;
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2. to support bone marrow recovery after radiation and chemotherapy;
3. to prevent secondary infections due to immunosuppression caused by standard chemotherapy and
surgical interventions;
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4. to increase complete and partial response rate to anticancer therapies; and
5. to improve lymphocyte function and biological defense mechanisms.
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I request that you protect my invention in the best possible way!
References
[i] Maurer, HR, Eckert, K, Stange, R: Einfluss der Therapie auf die antitumorale Immunotoxizität der
Leukozyten von Mamma-Tumorpatientinnen. Pers. Mitt. (1999).
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[ii] Mustacchi, G, Paves, L, Milani, S et al: High-dose folinic acid and fluouracil plus or minus
thymostimulin for the treatment of metastatic colorectal cancer: results of a randomised multicentered
trial. Anticancer Res. (1994) 14: 617-619.
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[iii] Schulof, RS, Loyd, MJ, Cleary, PA, et al: A randomized trial to evaluate the immunorestorative
properties of synthetic thymosin-alpha1 in patients with lung cancer. J Biol Resp Modif (1985) 4: 147158.
[iv] Azizi A, Brenner HJ, Shoham J: Postoperative adjuvante Behandlung von Patienten mit malignem
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Melanom durch den Bone marrowfaktor. Arzneim-Forsch/Drg Res (1984) 34(II): 1043-1046.
[v] Massimo F, Gobbi P, Moretti G, Avanzini P, Italian Lymphoma Study Group: Effects of
Thymostimulin with combination Chemotherapy in patients with aggressive non-Hogkins lymphoma.
Am J Clin Oncol (CCT) (1995) 18(1): 8-14.
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[vi] Peretti P, Tonelli F, Mazzei T, Ficari F, Italian study group on antimicrobal prophylaxis in
abdominal surgery. J Chemotherapy (1993) 5(1): 37-42.
[vii] Gonelli S, Petrioli R, Cepollaro C, Palmieri R, Aquino A, Gennari C: Thymostimulin in
association with chemotherapy in breast cancer patients with bone metastases. Clin Drug Invest (1995)
9(2): 79-87.
[viii] Iaffaioli RV, Frasci G, Tortora G, Ciardiello F, Nuzzo F, Scala S, Pacelli R, Bianco AR: Effect
of bone marrow extract on the incidence of infections and myelotoxicity during adjuvant
chemotherapy for breast cancer. Bone marrow (1988) 12: 69-75. Kluwer Academic Publishers.
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