CN101505781A - Lactoferrin as a radioprotective agent - Google Patents

Abstract

This present invention relates to the field of protecting against, or rectifying the effects of damaging ionizing irradiation. The method of treatment involves oral administration of a lactoferrin composition, alone or in combination with other treatments, both in combination with other radio-protective agents and/or the standard of care. Further, the method of treatment provides for a topical administration of lactoferrin to treat lesions caused by local damaging irradiation.

Description

Lactoferrin as radioprotective substance

Technical field

[0001] the present invention relates to medical domain, more specifically to the purposes of lactoferrin as radioprotective substance.Lactoferrin is used for preventing or repairing the effect of damaging ionizing radiation, and improves animals survived.

Background of invention

[0002] ionizing radiation mainly has a negative impact by cytotoxicity pair cell and tissue.In the mankind, the ionizing radiation that is subjected to mainly contacts generation by treatment technology (as antitumor radiotherapy) or by occupation with environment.

The main source of the ionizing radiation that [0003] is subjected to is the treatment radiation of using in cancer or the treatment of other proliferative disease.0.1 and 2 gray(Gy)s (Gy) are accepted in the each usually treatment of experimenter that is subjected to the therapeutic dose ionizing radiation, and can treat acceptance up to 5Gy at every turn.Depend on the therapeutic process of treatment doctor regulation, the experimenter can accept multiple dose in several weeks in the process of some months.

[0004] its work relates to the radiating people of contact (maybe may contact), and for example the people in nuclear energy and the nuclear weapon industry may be subjected to the ionizing radiation of occupation amount.At present 104 nuclear power plants that carry out commercial operation through permission are arranged in the U.S..In the world, one have 430 nuclear power plants 32 country's operations.The all staff on board who employs in these nuclear power plants specifies at it may the contact ionization radiation in responsibility process.The incident of Three Mile Island nuclear power plant on the 28th contingency March in 1979 for example, radioactive substance is discharged in containment vessel and the surrounding enviroment, shows potential harmful contact.Even without catastrophic event, the staff of nuclear power industry is subjected to higher levels of radiation than general public.

[0005] quarters at the army personnel that nuclear reactor drives boats and ships, or must the radiating risk of similar contact ionization be arranged the soldier of radioactive fallout IA operation.The occupational contact also may appear at by call treatment and relate among the rescue emergency worker of nuclear reactor and radioactive substance catastrophic event.For example the people of Chernobyl nuclear power plant on the 26th reactor fire fire fighting April in 1986 is subjected to radiation, and a lot of people die from radiation effect.In August, 2000, when naval and civilian post rescue personnel attempt the Russian nuclear-powered submarine of rescue sinking Kursk crewman, emitting the danger of raying.If the submarine reactor device damage, the salvor may still face radiation.

[0006] other sources of occupation contact can be machine components, plastic and the solvents that manufacturing is left over from the radioactivity medical product, smoke detector, emergency sign and other consumer goodss.Occupational contact also may occur in the personnel that hold office on the nuclear power ship, especially the army personnel who is subjected to the operation of nuclear weapon radioactive fallout Polluted area and handle guard's nuclear reactor among the Nuclear Accident Emergency personnel the people.

[0007] people and other animals (as domestic animal) also may contact the ionizing radiation from environment.The main source that contacts a large amount of environmental radiations is nuclear power plant's contingency, as the accident in Three Mile Island, Chernobyl and East Sea village.The research prediction of Sandia National Laboratory nineteen eighty-two, the nuclear accident of " worst case " can cause surpassing 100,000 death toll, and the long-term radioactive pollution in large tracts of land soil.

[0008] for example the estimation death toll of Chernobyl accident is 8,000 to 300,000, only is subjected in various degree radiation pollution in Ukraine with regard to the soil that surpasses 4,600,000 hectares is arranged.After the accident first week far away from Ireland, the Scandinavian Peninsula is northern and the littoral reflexive precipitum that just detects in Arras.135,000 people evacuate in " the dead district " of 30 miles of radiuses around the nuclear power plant of Chernobyl, and this area still is not suitable for human living now.Still the live area of " dead district " outer low-level radiation of about 1,200,000 people.

[0009] other nuclear power plant's accident has discharged a large amount of being radiated in the environment.Discussed above three miles island accidents.In Japan, the disruptive pipe leakage of in July, 1999 Tsuruga 2 nuclear power plants 51 tons of cooling waters.More serious accident occurs in the uranium treatment facility in JIUYUE in 1999 the Japanese East Sea on the 30th village, and 69 people are subjected to a large amount of radiation.When the workman has started self-holding nuclear chain reaction unintentionally accident having taken place, causes radiation to be discharged into the atmosphere.Detect radiation counter 0.84mSv/ hour in immediate zone (ultimate 4000 times of year).39 families (150 people) are evacuated, and are declared as the forbidden zone in 200 meters of the spot radiuses.Road in 3 kilometers of the spot radiuses is blocked, the resident in 10 kilometers of the spot radiuses be apprised of stay in indoor." criticality incident " in East Sea village on the nuclear power industry history, be positioned at the San Li village and and the Chernobyl after, be listed in the 3rd serious accident.

[0010] radiation of environment contact ionization also may be exploded from nuclear weapon (no matter being experimental or durante bello), and nuke rubbish stores and the acitinide of nuclear fuel processing and reprocessing discharges, and spontaneous radioactive substance such as radon gas or uranium.The ammunition that use contains depleted uranium causes the low-level radioactive pollution in area of hostilities, also causes more and more the concern.

[0011] skin time-delay, irreversible variation, radiation dermantitis or radiodermatitis be not the inferior total irradiation that causes death usually, but be limited to skin more high-dose irradiation caused.These variations may take place, if the heavily contaminated of beta emitter to baring skin for example arranged.Table 4 has been listed the radiation dermantitis degree of local skin zone radiological dose.

Table 4: radiation dermantitis

[0012] radiation induced damage can be repaired, and is coarse but repair in some cases, the late effect that caused after adverse health influence in a few hours or short time several weeks or the radiation a lot of months or year can observe.Radiation induced germinal mutation can cause the hereditary change that many generations may not express.The performance of adverse health influence is depended in radiation dose certainly, is contacted radiating time, histological difference and sensitivity and inherent anti-oxidative defense mechanism.

[0013] ionizing radiation can make immune system depletion or be suppressed.Most of suppress to be attributable to radiation or albumen destroy due to cell death or the functional disorder cell injury or the death that directly cause, DNA is synthetic to suppress the DNA that causes or RNA chain interruption or the like.Press for and differentiate prevention and, be used to have the personnel and the harmful ionizing radiation personnel's of contact of contact radiation risk treatment from the non-toxicant of healing of radiation damage.

[0014] acute effect of high dose radiation comprises hematopoietic cell loss, immunosuppressant, mucosa (gastrointestinal tract and mouth) damage, and may damage other positions such as lung, kidney and central nervous system.The long term of high dose and low dose radiation consequence comprises the dysfunction or the fibrosis of many organs and tissue, and cancer.These variations are reflected on quality of life and the human mortality.

[0015] infecting is the lethal main cause of ionizing radiation dose that causes hemopoietic and GI symptom.Follow the high dose radiation of GI damage to cause antibacterial from intestinal other position migration and increase mortality rate in body.

[0016] the medicine radioprotectant provides cost efficient, radioprotector alternative effective and that obtain easily.Yet pharmaceutical composition attempted all not becoming fully merits and demerits in Normocellular radiation protection in the past.For example, when giving before the radiation, bone marrow protection effect (Neta et al., Semin.Radiat.Oncol.6:306-320,1996) is arranged, but do not have the system protection effect at the mobilization peripheral blood progenitor cells factor.Other chemical radiation protective agent is used separately or is share the less protective effect that shows mice with biological response modifier; but it is these compound administration are not too successful when the large mammal; thereby whether the chemical radiation protection has any value to be subjected to query (Maisin; J.R.; Bacq andAlexander Award Lecture. ＂ Chemical radioprotection:past; present; and future prospects ＂; Int J.Radiat Biol.73:443-50,1998).The medicine radiosensitizer that known preferential enhanced rad acts in cancerous tissue obviously is not suitable for the General System protection of contact ionization radiation normal structure.

[0017] adverse effect that is free radical because of radiation-induced cell injury main cause, the molecule with direct removing free radical performance especially is hopeful as radiation protective.The most famous radiation protective is a sulfhydryl compound, as cysteine and mercaptoethylmaine.But these chemical compounds produce serious adverse, as feeling sick, vomitting, are considered to poisonous under needs radiation protection dosage.Amifostine (WR-2721) is though to be used for radiotherapy through the approval of food and Drug Administration clinical, carry for the reply solar flare when report U.S. spaceman moon trip is also arranged, must rescue and evacuate active emergency personnel but amifostine has the overview of side effect that it is not suitable for.Side effect comprises hypotension, feels sick, vomiting, sneeze, hectic fever, slight drowsiness and hypocalcemia, and serious in being enough to limit the required drug dose that is lower than needs realization greatest irradiation protective effect level.In addition, amifostine have only when intravenous injection (iv) or be effectively during subcutaneous injection (SC), so its actual administration is difficult, the effectiveness in the act of terrorism (open-field terrorism) is low especially out of doors.Another radioprotective substance Cystapos (WR-638) is effective when having only intravenously administrable.Another compound d-CON (WR-1607) or raticide (sudden cardiac arrest causes death), as if more effective than amifostine, can produce equal protective effect with 1/100 described dosage.Yet, similar with amifostine, because its extreme toxicity d-CON is found and can't uses.Though another material promotes the male estradiol of hemopoietic system to propose the prophylactic thing as ionizing radiation-resistant, only estimates in laboratory animal so far.Potassium iodide (IOSAT ^TMKI) be that unique Food and Drug Administration ratifies, the thyroid block medicine of paper tinsel sealing is used for preventing that the people who is exposed to radioiodine in radiation emergency process from suffering from thyroid carcinoma.This medicine has been advised not only the Emergency Planning Zone at 10 miles, and may use by affected area at any or all.KI makes thyroid saturated with stable iodine, absorbs radioiodine thereby hindered thyroid.But the radioiodine that KI takes precautions against is the side-product (in the disaster process of Chernobyl) of the nuclear fission that only takes place in nuclear reactor, may not can occur in " dirty bomb " explodes, and has limited the application of potassium iodide.

[0018] though write down anti-low linear energy transfer (representing) with KeV/ μ m, the performance of most of known radioprotectants of the damage that causes as gamma ray and X ray (from 0.2 to 2.0KeV/ μ m) ionizing radiation, but their anti-high linear energy transfer radiation such as proton, neutron and alpha-particle (from 4.7 to 150KeV/ μ m) effectiveness of the damage that causes, in occurring in the nuclear device blast, also thoroughly do not studied.

[0019] melatonin may be applied in this and is worth mentioning as ionizing radiation-resistant is protectant.Melatonin is effectively removed hydroxyl and other free radicals, together with its indirect antioxygenic property, writes down (more than 900 kind of publication in the document) repeatedly in many independently investigation.The animal that is subjected to total irradiation and gives melatonin shows survival increases (LD _50/30And fatal radiation dose); The inductive oxidative damage protection of radioprotective also is tangible in hemopoietic but also at its hetero-organization not only.The more important thing is that different with amifostine, oral melatonin produces the higher cyclical level and the tissue concentration of faster increase.

[0020] therefore determine novel, nontoxic, effectively and easily chemical compound is avoided the damaging action of ionizing radiation to protect the mankind, remain to press for.The present invention at first uses the oral lactoferrin compositions, prevention or the somatic damage that causes of treatment ionizing radiation, and improve patient's existence.

The invention summary

[0021] the present invention relates to somatic damage preventative or that the radiation of therapeutic treatment contact ionization causes, and improve the method for patient's existence.This Therapeutic Method comprises oral (for example other radioprotective substance) the oral lactoferrin compositions of share separately or with other treatment.In another embodiment, the lactoferrin that occurs in the topical preparation is used for the treatment of the dermatosis that the local damage radiation causes.

[0022] as described herein, the easier definition the present invention of the sentence of following column number.

1. treatment contacts radiating experimenter's method, and it comprises the step that the lactoferrin compositions of effective dose is applied to the experimenter, and wherein said lactoferrin compositions reduces sickness rate and/or the mortality rate that is subjected to the radiation experimenter.

2. sentence 1 described method, described lactoferrin compositions is used before being subjected to radiation.

3. sentence 1 described method, described lactoferrin compositions is used after being subjected to radiation.

4. sentence 1 described method, wherein said lactoferrin compositions is dispersed in the pharmaceutically acceptable carrier.

5. sentence 1 described method, the amount of the described lactoferrin compositions of wherein using is about 0.01 to 2.0g/kg every day.

6. sentence 1 described method, the amount of the described lactoferrin compositions of wherein using is about 0.01 to 0.5g/kg every day.

7. sentence 1 described method, wherein said lactoferrin compositions oral administration.

8. sentence 7 described methods, wherein said lactoferrin compositions is with the liquid preparation administration.

9. sentence 7 described methods, wherein said lactoferrin compositions is with the solid preparation administration.

10. sentence 9 described methods, wherein said solid preparation comprises enteric coating.

11. sentence 1 described method, wherein said lactoferrin compositions topical.

12. sentence 1 described method, wherein said radiation is selected from ²³⁵U, ¹³¹I, ¹²³I, ⁹⁹Tc, ²⁰¹Th, ¹³³Xe, ¹²⁵I, ⁶⁰Co and ¹³⁷Cs, ⁶⁰Co, ¹³⁷Cs, ¹⁹²Ir, ³²P, ⁹⁰Sr, ²²⁶Ra and combination thereof.

13. treatment is exposed to the method for the sequela that the doses ionizing radiation caused, it comprises the step of replenishing experimenter's mucomembranous immune system by orally give effective dose lactoferrin compositions.

14. strengthen the method for the gastrointestinal tract mucous immunne response of experimenter that is subjected to the absorbed dose ionizing radiation, it comprises the step of orally give effective dose lactoferrin compositions.

15. sentence 14 described methods, the generation of wherein said lactoferrin compositions stimulating cytokine or chemotactic factor.

16. sentence 14 described methods, wherein said lactoferrin compositions causes the inhibition of cytokine or chemotactic factor.

17. sentence 15 described methods, wherein said cytokine are selected from interleukin-18 (IL-18), interleukin 12 (IL-12), granulocyte/M-CSF (GM-CSF) and IFN-(IFN-γ).

18. sentence 15 described methods, wherein said chemotactic factor are macrophage inflammatory protein 3 α (MIP-3 α), macrophage inflammatory protein 1 α (MIP-1 α), macrophage inflammatory protein 1 β (MIP-1 β).

19. sentence 16 described methods, wherein said cytokine are selected from interleukin II (IL-2), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 10 (IL-10), and tumor necrosis factor-alpha (TNF-α).

20. sentence 33 described methods, wherein said lactoferrin compositions suppresses the generation of matrix metalloproteinase (MMPs).

21. sentence 17 described methods, the wherein generation or the activity of interleukin-18 or granulocyte/M-CSF immune stimulatory cell.

22. sentence 21 described methods, wherein said immunocyte is selected from T lymphocyte, natural killer cell, macrophage, dendritic cell and polymorphonuclear cell.

23. sentence 22 described methods, wherein said polymorphonuclear cell is a neutrophil.

24. sentence 22 described methods, wherein said T lymphocyte is selected from CD4+, CD8+ and CD3+T cell.

25. reduce the method for the experimenter's mortality rate that is subjected to the absorbed dose ionizing radiation, it comprises the described experimenter of lactoferrin compositions orally give with effective dose, to alleviate the step of described absorbed dose effect.

26. alleviate the method for absorbed dose radiation to experimenter's damaging action, it comprises the described study subject effective dose of orally give lactoferrin compositions, to alleviate the step of described absorbed dose damaging action.

27. sentence 26 described methods wherein alleviate described damage and cause described experimenter's mortality rate to reduce.

28. sentence 26 described methods wherein alleviate described damage and cause intestinal related system antibacterial, virus or fungal infection to reduce.

29. sentence 26 described methods wherein alleviate described damage and cause described experimenter's mortality rate to reduce.

30. alleviate the method for absorbed dose radiation to experimenter's damaging action, it comprises orally give described experimenter's effective dose lactoferrin compositions and radioprotective substance, with the step of the damaging action that alleviates described absorbed dose.

31. sentence 30 described methods, wherein said radioprotective substance are granulocyte stimulating factor (G-CSF) (filgrastim/(Neupogen)) or amifostine.

32. treatment is subjected to the method for the sequela that the doses ionizing radiation causes, it comprises the step of replenishing experimenter's mucomembranous immune system by the lactoferrin compositions of local application effective dose.

Be subjected to the method that the absorbed dose ionizing radiation causes experimenter's skin histology immunne response of radiation dermantitis 33. strengthen, it comprises the step of local application effective dose lactoferrin compositions.

34. sentence 33 described methods, the generation of wherein said lactoferrin compositions stimulating cytokine or chemotactic factor.

35. sentence 33 described methods, wherein said lactoferrin compositions causes the inhibition of cytokine or chemotactic factor.

36. sentence 35 described methods, wherein said cytokine are selected from interleukin-18 (IL-18), interleukin n-12 (IL-12), granulocyte/M-CSF (GM-CSF) and IFN-(IFN-γ).

37. sentence 35 described methods, wherein said chemotactic factor are macrophage inflammatory protein 3 α (MIP-3 α), macrophage inflammatory protein 1 α (MIP-1 α), macrophage inflammatory protein 1 β (MIP-1 β).

38. sentence 35 described methods, wherein said cytokine are selected from interleukin II (IL-2), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 10 (IL-10), and tumor necrosis factor-alpha (TNF-α).

39. sentence 33 described methods, wherein said lactoferrin compositions suppress matrix metalloproteinase (MMPs) and produce.

40. sentence 36 described methods, the wherein generation or the activity of interleukin-18 or granulocyte/M-CSF immune stimulatory cell.

41. sentence 40 described methods, wherein said immunocyte is selected from T lymphocyte, natural killer cell, macrophage, dendritic cell and polymorphonuclear cell.

42. sentence 41 described methods, wherein said polymorphonuclear cell is a neutrophil.

43. sentence 41 described methods, wherein said T lymphocyte is selected from CD4+, CD8+ and CD3+T cell.

[0023] characteristics of the present invention and technical advantage have been summarized more than quite widely, so that detailed description of the present invention afterwards can be better understood.Below will describe other characteristics of the present invention and advantage, it has formed the theme of claim of the present invention.It will be understood by those skilled in the art that disclosed notion and specific embodiments are easy of making an amendment or designing other structures to realize the basis of the identical purpose of the present invention.Those skilled in the art it should further be appreciated that the structure of this equivalence does not deviate from the spirit and scope of the present invention as stating in the accessory claim.When considering with accompanying drawing, from following description, be considered to the distinctive new feature of the present invention about its structure and operational approach, will be better understood with further theme and advantage.But should understand clearly, each figure only provides for illustrating and describing purpose, is not to be defined as limitation of the present invention.

The accompanying drawing summary

[0024] in order more completely to understand the present invention, with reference now to following explanation and accompanying drawing, wherein:

[0025] Fig. 1 is the mice survival rate that is subjected to the ionizing radiation of about 10Gy whole body fatal dose.Dotted line is represented the mice that his lactoferrin is handled, and solid line is represented the placebo mice.

[0025] Fig. 2: handle the recovery of the circulation medium-sized lymphocyte that quickens radiation exhaustion with his lactoferrin.Before showing radiation among the figure, and the FACS of each time point leukocyte sum behind the non-deadly radiation murine of about 5Gy whole body.* represent non-matching, two tail P value 0.0359.

[0025] Fig. 3 is a mice health status score value after the 6Gy radiation.This data set N=20, placebo has remarkable different end point values (p=0.0259) with his lactoferrin cohort.

Detailed Description Of The Invention

[0026] the disclosed the present invention of the application can have various embodiments and distortion, and is not inclined to one side From scope and spirit of the present invention, this is apparent to those skilled in the art .

I. definition

[0027] just as used herein, word " " and claim and/or specification In term " comprise " when using together, can refer to " one ", but can also with " one or many Individual ", the meaning of " at least one " and " or above " is consistent. Further come Say that term " has ", " comprising ", " containing " and " comprising " can exchange art technology people It is open-ended term that the member has known these terms.

[0028] term used herein " lactoferrin compositions " refer to that lactoferrin is arranged, breast The part of ferritin or part, the terminal Lactoferrtin variants of N-or its combination.

[0029] term used herein " lactoferrin " or " LF " refer to natural or recombinate newborn iron Albumen. The genuine milk ferritin can be from mammal milk or colostrum or natural next from other The source purifying obtains. Recombinant lactoferrin (rLF) can pass through transgenic animals, plant, Recombinant expressed or the direct preparation of fungi, bacterium or other protokaryons or eucaryon species, or pass through Chemical synthesis makes.

[0030] term used herein " human lactoferrin " or " hLF " refer to natural or recombinant human Lactoferrin. The natural human lactoferrin can be from people's milk or colostrum or natural from other The source purifying obtains. Restructuring lactoferrin (rhLF) can be by transgenic animals, plant Recombinant expressed or the direct preparation of thing, fungi, bacterium or other protokaryons or eucaryon species, or Make by chemical synthesis.

[0031] term used herein " bovine lactoferrin " or " bLF " refer to natural or recombinant bovine Lactoferrin. Natural bovine lactoferrin can obtain from cow's milk juice purifying. Restructuring cow's milk iron Albumen (rbLF) can pass through transgenic animals, plant, fungi, bacterium or other protokaryons Or the recombinant expressed or direct preparation of eucaryon species, or make by chemical synthesis.

[0032] term used herein " the terminal Lactoferrtin variants of N-" refers to newborn iron egg In vain, wherein at least N end glycine by brachymemma and/or replacement. The terminal lactoferrin of N-becomes Body also includes but not limited to delete and/or replace one or more-terminal amino acid residues, For example 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15 or 16-terminal amino acid residues etc. Therefore, the terminal Lactoferrtin variants of N-comprises At least 1 to 16-terminal amino acid residue disappearance or brachymemma and/or replacement. At least N-Terminal glycine disappearance and/or the lactoferrin that replaces are regulated same with the total length lactoferrin Biological effect, and/or can strengthen the biologically active of lactoferrin, for example by stimulating each Plant the generation (for example IL-18, MIP-3 α, GM-CSF or IFN-γ) of cell factor, logical Cross and suppress various cell factors (for example IL-2, IL-4, IL-5, IL-10 or TNF-α), and by improving other promotions or strengthening the parameter of experimenter's health status.

[0033] term used herein " oral administration " includes but not limited to oral, mouth Chamber, enteron aisle or intragastric administration.

[0034] term used herein " immunity weakens " is defined as when the potential disease of contact During substance, because one or more anti-infective normal defense mechanisms reductions of experimenter, and not Can be fully and effectively cause of disease is made the experimenter's who replys state, such state is Caused by the ionising radiation of described sustaining damage property of experimenter type and dosage. Surpass one The defective of planting physical mechanism may be affected (such as bone marrow injury; Immune blood The exhaustion of lymphocyte, BMDC and other cells; Damage and permeability thereupon Increase, thereby epithelium (such as intestines, skin, lung) protective effect reduces etc.).

[0035] described " state that immunity weakens " used herein is Body contact or agent Amount absorbs the consequence of all kinds and strength damage ionising radiation. Ionising radiation is particle Radiation, wherein single particle (for example photon, electronics or helion) carries enough energy Make atom or molecular ionization (namely removing electronics fully from its track). If these ionization Occur abundantly, can very strong destructiveness be arranged to living tissue. The composition of ionising radiation can become Change. If the energy of each photon is high (being that wavelength is enough short) enough, then electromagnetic radiation Can cause ionization. Far ultraviolet, X ray and gamma rays all are ionising radiations. Ionization Radiation also can by the particle of fast moving, form such as electronics, positive electron or small nuclei.

Term used herein " parenteral " comprises any type of administration, wherein institute Stating compound absorbs not relate in the subject and passes through intestinal absorption. Be used for of the present invention Typical case's parenteral includes but not limited in intramuscular injection, intravenous injection, the peritonaeum, eye In or intra-articular administration.

[0036] term used herein " pharmaceutically acceptable carrier " comprises any and institute Have solvent, decentralized medium, dressing, antibiotic and antifungal agent, etc. blend the delay absorbent Etc.. Medium and the material such for the material use of pharmaceutical active are many in this area Known. Unless arrived any conventional media or material and carrier of the present invention or cell Incompatible degree, otherwise consider its use in pharmaceutical composition. The work that replenishes The property composition also can be impregnated in the described composition.

[0037] term used herein " pharmaceutical composition " refers to be dispersed in pharmacy and can accept to carry Lactoferrin compositions in the body. Described lactoferrin compositions can comprise lactoferrin or N holds Lactoferrtin variants, wherein at least the terminal glycine amino acid residue of N-by brachymemma or Replace.

[0038] term used herein " experimenter " refers to according to method mouth described herein Clothes give any mammalian subject of human lactoferrin composition. In implementation side In the case, adopt method treatment human experimenter of the present invention.

[0039] term used herein " treatment effective dose " is instigated disease symptoms to improve or is mended The amount of rescuing.

[0040] term used herein " topical " includes but not limited to part, skin (as in corium or corium), epidermis or subcutaneous.

[0041] term used herein " treatment " refers to the recombinant human lactoferrin with the treatment effective dose Protein composition is applied to the experimenter, thereby experimenter's disease takes a turn for the better. Describedly transfer well disease to Any improvement of shape or remedy. Described improvement is Observable or measurable improvement. Cause This, those skilled in the art recognize that treatment may improve described disease but are may not can complete The described disease of full healing.

II. lactoferrin

[0042] lactoferrin used according to the invention can by separate from natural origin and Purifying obtains, such as but not limited to mammal milk. The lactoferrin preferred mammal Lactoferrin is such as ox or human lactoferrin. In preferred embodiments, described newborn iron Albumen is with technique for gene engineering restructuring preparation well known and that use, and is moving such as transgenosis Thing, plant or Eukaryotic recombinant expressed or direct preparation, or chemical synthesis. Referring to For example U.S. Patent number 5,571, and 896,5,571,697 and 5,571,691, it draws work at this Reference.

[0043] in some aspects, the invention provides and have natural LF and/or rLF The natural activity that strengthens is as stimulating and/or suppress the ability of cell factor or chemotactic factor (CF) Lactoferrtin variants. In particular, the invention provides at least the terminal glycine residue of N-by The Lactoferrtin variants of replacement and/or brachymemma. The terminal Lactoferrtin variants of N-may be natural Produce, or modified by one or more 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factors or disappearance.

[0044] United States Patent (USP) 6,333,311 that described disappearance variant can be incorporated by reference such as this paper Described, by the multinuclear glycosides of lactoferrin of lactoferrin proteolysis and/or coding brachymemma The expression of acid is prepared.

[0045] replacement variant or replacement variant contain in the described albumen one or more usually Another amino acid of amino acid substitution in site. Replacement can be guarded, just Say that an amino acid is with the 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor of an analogous shape and electric charge. Conservative being substituted in This area is well-known, and for example comprise: alanine becomes serine; Arginine becomes Be lysine; Asparagine becomes glutamine or histidine; Aspartic acid becomes paddy ammonia Acid; Cysteine becomes serine; Glutamine becomes asparagine; Glutamic acid becomes Aspartic acid; Glycine becomes proline; Histidine becomes asparagine or glutamy Amine; Isoleucine becomes leucine or valine; Leucine becomes valine or different bright ammonia Acid; Lysine becomes arginine; Methionine becomes leucine or isoleucine; Phenylpropyl alcohol ammonia Acid becomes tyrosine; Leucine becomes methionine; Serine becomes threonine; Threonine Become serine; Tryptophan becomes tyrosine; Tyrosine becomes tryptophan or phenylpropyl alcohol ammonia Acid; Valine becomes isoleucine or leucine.

[0046] in making such change, but the hydrophilic index of considered amino acid. Compose Give the importance of hydrophile amino acid index in the biological function of protein-interacting in this area extensively understood (Kyte and Doolittle, 1982). Admittedly, amino acid whose phase Hydrophilic nmature is helped thereby the secondary structure of the albumen that obtains, and this determines again conversely Albumen and other molecules such as enzyme, substrate, acceptor, DNA, antibody, antigen etc. Interact.

[0047] each has amino acid basedly specified hydrophilic finger in its hydrophobicity and charge characteristic Number (Kyte and Doolittle, 1982), these are: isoleucine (+4.5); Valine (+4.2); Leucine (+3.8); Phenylalanine (+2.8); Cysteine/cystine (+2.5); Methionine (+1.9); Alanine (+1.8); Glycine (0.4); Threonine (0.7); Silk ammonia Acid (0.8); Tryptophan (0.9); Tyrosine (1.3); Proline (1.6); Histidine ( 3.2); Glutamic acid (3.5); Glutamine (3.5); Aspartic acid (3.5); Asparagine ( 3.5); Lysine (3.9); And arginine (4.5).

[0048] some amino acid known in the art can with other have similar hydrophilic index or The 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor of value is still so that protein has similar biologically active, namely still Obtain the suitable protein of biological function. In making this change, preferred index of affinity 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor in ± 2 scopes, especially preferred those in ± 1 scope, even More preferably those in ± 0.5 scope.

[0049] this area is also understood, and can effectively carry out similar ammonia on hydrophilic basis The replacement of base acid. The United States Patent (USP) 4,554,101 that is hereby incorporated by reference is illustrated, and is closed on by it The biology of the maximum local average hydrophily of the albumen that the amino acid pro water-based determines and described albumen Characteristic is relevant. As United States Patent (USP) 4,554,101 detailed descriptions, amino acid residue is referred to Fixed following hydrophilicity value: arginine (+3.0); Lysine (+3.0); Aspartic acid (+3.0 ± 1); Glutamic acid (+3.0 ± 1); Serine (+0.3); Asparagine (+0.2); Glutamine (+0.2); Glycine (0); Threonine (0.4); Proline (0.5 ± 1); Alanine ( 0.5); Histidine (0.5); Cysteine (1.0); Methionine (1.3); Valine ( 1.5); Leucine (1.8); Isoleucine (1.8); Tyrosine (2.3); Phenylalanine ( 2.5); Tryptophan (3.4).

[0050] further, another has similar parent to be understood that the available amino end acid substitution The amino acid of water number still obtains the quite protein suitable with immunology of biology. At this In the variation of sample, preferably the 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor of its hydrophilicity value in ± 2 scopes is especially preferred In ± 1 scope those, even more preferably those in ± 0.5 scope.

[0051] therefore, in the present invention, but replace variant or the mutagenesis of replacement Application standard Technology, for example United States Patent (USP) 5,220,007,5,284,760,5,354,670, Open in 5,366,878,5,389,514,5,635,377,5,789,166 and 6,333,311 Rite-directed mutagenesis preparation, described patent is hereby incorporated by reference. Envision at least terminal sweet ammonia of N-The acid residue can be used amino acid substitution or the replacement of arbitrarily 20 natural generations, for example just Electric charge amino acid (arginine, lysine or histidine), neutral amino acid (third ammonia Acid, asparagine, cysteine, glutamine, glycine, isoleucine, bright ammonia Acid, methionine, phenylalanine, proline, serine, threonine, tryptophan, junket Propylhomoserin, valine) and/or the amino acid (aspartic acid and glutamic acid) of negative electrical charge. Advance One step ground, imagining amino acid residue in any N1 to N16 scope can be replaced or get Generation. Anticipation then reaches most at least as long as described albumen keeps its biology and/or functional activity 16-terminal amino acid residues can be replaced or be replaced, by suppress various cells because of The son (such as IL-2, IL-4, IL-5, IL-10 and TNF-α) and/or with regard to its medical conditions and Speech by improving and promoting or strengthen the healthy relevant parameter of experimenter, stimulates various thin The generation of intracellular cytokine (such as IL-18, MIP-3 α, GM-CSF or IFN-γ). Therefore, The terminal Lactoferrtin variants of N-of the present invention is considered to the function equivalent of lactoferrin.

[0052] about function equivalent, those of skill in the art understand " biological function well Be equal to " albumen definition in inherent concept be: when the life that is equal to that keeps the molecule acceptable level Thing is active and/or when strengthening the biologically active of described lactoferrin molecule, at described molecule Can make the restricted number of change in the determining section. Therefore the biological function equivalent is at this paper Being defined as wherein selecting amino acid (or codon) may substituted those egg In vain. Functional activity is defined as the lactoferrin stimulation or suppresses various cell factors or chemotactic The factor, and/or with regard to its medical conditions, promote or strengthen experimenter's health by improving The ability of parameter. For instance, prolong experimenter's any a period of time of life; Reduction The infringement that radiation causes; Accelerate the normalization of experimenter's compromised immune system; Minimizing can be returned Cause is in experimenter's pain of experimenter's disease.

[0053] further, described-terminal amino acid residue can with modify and/ Or rare 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor. This paper hereinafter provides typical but nonrestrictive modification And/or rare amino acid whose table.

[0054] the terminal lactoferrin of N-in lactoferrin (lactoferrin compositions) preparation The existence of variant (disappearance and/or replacement) and relative scale can be passed through the Application standard method The Edman degradation process is measured the-terminal amino acid sequence and is determined. The terminal lactoferrin of N-The relative scale of variant comprises at least 1% lactoferrin compositions, at least 5% lactoferrin Composition, at least 10% lactoferrin compositions, at least 25% lactoferrin combination Thing, at least 50% lactoferrin compositions or between any scope.

[0055] in the method, described protein and phenyl isothiocyanate (PITC) are anti-Should, PITC reacts with aminoterminal amino acid residue under alkali condition, forms phenylamino Base formyl sulfide radical derivative (PTC-albumen). Trifluoroacetic acid first amino that ruptures then Acid forms its Dimazole quinoline ketone (anilinothialinone) derivative (ATZ-amino Acid), stay new amino terminal and carry out next round degraded circulation.

[0056] percentage of the terminal Lactoferrtin variants of N-can also be by using red sulphonyl Changing reaction (Dansylation reaction) determines more accurately. Briefly, alkaline bar (pH10) use makes the red sulfonylation of albumen with the dansyl Cl of described albumino reaction under the part. Red Behind the sulfonylation, reactant mixture is dried to particle, then complete water in 6N hydrochloric acid Separate.-terminal amino acid uses fluorescence photometer in the line by RP HPLC, and uses known pellet The standard of sulfonyl amino acid preparation is relatively differentiated.

III. pharmaceutical composition

[0057] the present invention submits to comprise the lactoferrin combination that is dispersed in the pharmaceutical carrier to Thing. The lactoferrin that contains in the composition of the present invention comprises lactoferrin or the terminal breast of N-The ferritin variant, wherein at least the terminal glycine residue of N-1 by brachymemma or replacement. The N-end The end Lactoferrtin variants comprise lack at least the terminal glycine residue of N-or contain the N-end sweet The variant that the propylhomoserin residue replaces. Described replacement can comprise with nature or artificial amino acid Residue replaces the terminal glycine residue of described N-. For example, described replacement can comprise with just Electric charge amino acid residue or negative electrical charge amino acid residue replace the terminal glycine residue of N-, or Replace the terminal glycine residue of N-with the neutral amino acid residue except glycine. Other N-ends The end Lactoferrtin variants comprises and lacks one or more N-terminal residues or have one or more The terminal lactoferrin that replaces of N-. The terminal Lactoferrtin variants of N-comprises at least 1% group Compound, at least 5% composition, at least 10% composition, at least 25% combination Thing, at least 50% composition or between any scope.

[0058] further according to the present invention, the present composition that is suitable for administration is having Or do not have to provide in the pharmaceutically acceptable carrier of inert diluent. Described carrier should be can Assimilation, comprise liquid, semisolid, such as cream or solid carrier. Unless any conventional Jie Matter, material, diluent or carrier are unfavorable for the recipient or the controlling of the composition that wherein contains Therapeutic effect, it is suitable using in the composition purposes that is used for putting into practice the inventive method . The example of carrier or diluent comprise fat, oil, water, physiological saline, lipid, Liposome, resin, adhesive, filler etc., or its combination.

[0059] according to the present invention, described composition and described carrier are with any convenience and reality With mode make up, for example by dissolving, suspendible, emulsification, mixing, encapsulation, absorption Deng. Such program is conventional to those skilled in the art.

[0060] in specific embodiments of the present invention, described composition with semi-solid or Solid carrier thoroughly merges or mixes. Mixing can any easily mode be advanced such as grinding OK. Stabilizing agent can also add in mixed process, does not lose treatment with the protection group compound Activity, for example sex change in the stomach. The example that is used for described composition stabilizing agent comprises buffering Liquid, amino acid such as glycine and lysine, carbohydrate such as glucose, mannose, Galactolipin, fructose, lactose, sucrose, maltose, sorbierite, sweet mellow wine etc., albumen Hydrolase inhibitor etc. The combination of the oral administration of being combined with semisolid or solid carrier Thing can further be mixed with hard or soft shell capsule, tablet or pill. More preferably, glue Capsule, tablet or pill are by enteric coating. Enteric coating prevent composition the stomach of pH acidity or The upper intestines sex change. Referring to for example U.S. Patent number 5,629,001. In case the arrival small intestine, Alkaline pH value dissolving dressing wherein discharges lactoferrin compositions, and by special Cell uptake, such as enterocyte and Peyer ' s patch M cell.

[0061] in another embodiment, make pulverous composition and be with or without The liquid-carrier of stabilizing agent is such as water or physiological saline combination. The embodiment of introduction can Comprise the preparation excipient, such as carbomer, PEG, anticorrisive agent etc.

[0062] composition of the present invention can be mixed with neutrality or salt form. Pharmaceutically can connect The salt that is subjected to comprises acid-addition salts (forming with the free amine group group of albumen), acid-addition salts Be and inorganic acid example hydrochloric acid or phosphoamino acid, or such as acetic acid, oxalic acid, tartaric acid, flat The organic acid of peach acid etc. forms. With the salt that the free carboxy group forms also can be from nothing Machine alkali, the hydroxide of sodium, potassium, ammonium, calcium or iron for example, and such as isopropylamine, three The organic base of methylamine, histidine, procaine etc.

[0063] can pass through common approaches according to lactoferrin compositions of the present invention, mouthful Head, stomach and intestine are outer or topical. Typical case's approach comprises but not in oral, nasal cavity, mouth Chamber, rectum, vagina, stomach and intestine are outer, in the intramuscular, peritonaeum, in the intravenous, artery or skin Skin. Such composition is usually as described herein, gives as the acceptable composition of pharmacy Medicine.

The amount of the lactoferrin of [0064] using among the present invention may change, about 0.01 to 2.0g/kg preferred 0.01 to 0.5g/kg, single dose or divided dose. In preferred embodiment In, composition of the present invention comprises concentration approximately in solid, semisolid or liquid formulations 0.1% to about 100% lactoferrin. Lactoferrin compositions may comprise lactoferrin Or the terminal Lactoferrtin variants of N-, wherein the terminal glycine residue of at least described N-1 is cut Weak point and/or replacement.

[0065] once preparation, solution is in the mode compatible with described dosage particles, and with The such amount administration that effectively causes doing well,improving in the treatment or remedy. Described preparation is with each Plant dosage form, easy such as absorbable solution, medicament slow release capsule, barrier cream etc. The ground administration. Dosage can depend on that the experimenter's who is treated with quilt situation produces some variations. Negative The people of duty administration can under any circumstance, determine single experimenter's suitable dose. This Outward, for to people's administration, preparation need satisfy the FDA biological product standards and do (FDA Office of Biologics standards) aseptic, the common safety and the purity mark that require Accurate.

IV. treatment

[0066] according to the present invention, oral or be locally applied to suspect or be subjected to radiating experimenter with the lactoferrin compositions that provides in any above-mentioned pharmaceutical carriers, or before being subjected to radiation, be applied to the experimenter.Those skilled in the art can consider that as absorption, metabolism, medication, age, body weight, the ionization damage order of severity and the reaction to treating, decision is applied to the treatment effective dose of experimenter's lactoferrin based on some.The oral administration of lactoferrin compositions comprises mouth, oral cavity, enteral or intragastric administration.Also imagine described compositions and can be used as food additive.Be sprinkled upon on the food before for example described compositions is taken in or be added in the liquid.The topical of lactoferrin compositions comprises part, skin, epidermis or subcutaneous administration.

[0067] therapeutic scheme also can change, and usually depends on ionization damage or radiating type, damages or contact radiating position, the progression of disease that damage or contact radiation cause, patient's health status and age.Obviously the disease of some type will need more positive therapeutic, and some patient then is impatient at the more scheme of effort simultaneously.Based on the treatment known curative effect of preparation and toxicity (if any words), the clinicist will be best suited for making such decision.

[0068] described lactoferrin compositions can be used as the solution in the suitable buffer agent, or the solid oral dosage of capsule, tablet or similar suitable form, or as the topical formulations oral administration.The amount of the lactoferrin of using is 0.01 to 2.0g/kg, and preferred 0.01 to 1.0g/kg, single dose or divided dose.Imagination treatment lasts till and damages normalization, preferred 30 days continuous treatment.Therapeutic effect can be by measuring the composition of peripheral blood cells, and especially the content of leukocyte in the circulation is more generally monitored by experimenter's whole health.

[0069] treatment may comprise various " unit dose ".Unit dose be meant contain calculate to produce expectation with lactoferrin compositions administration, the i.e. suitable amount of pre-determining of the relevant therapeutic combination of replying of approach (lactoferrin compositions) with therapeutic scheme.The amount of using, and particular approach and preparation are in clinical technology personnel's skill.It is also important that experimenter to be treated, especially experimenter's state and desired protection.The unit dose of intravenous injection or subcutaneous injection administration does not need the single injection administration, but can comprise the continuous injection of a period of time.

[0070] in specific embodiments, lactoferrin compositions provides with single dose or multiple dose.But single dose administration every day, or inferior more than a day, and perhaps a week is repeatedly.In further embodiment, described lactoferrin compositions gives with a series of dosage.But the dosage every day of described series, or more than a day time, a week repeatedly or one week multiple dosing.

[0071] in a preferred embodiment of the invention, lactoferrin compositions is used with effective dose, preventing, reduce, to reduce or to suppress the damage that the health radiation of damaging ionizing radiation causes, and improves patient's survival rate.The amount of the lactoferrin of using is 0.01 to 2.0g/kg, and preferred 0.01 to 0.5g/kg, single dose or divided dose.The treatment imagination lasts till damages normalization, preferred 30 days continuous treatment.

[0072] improves any observable or measurable variation that is meant improvement.Described compositions of the present invention and method can reduce the radiating experimenter's of sustaining damage property mortality rate.In other respects, compositions of the present invention is used with effective dose, and reduce, reduce, suppress, prevent or eliminate immune system cell and damage and afunction, and the afunction of the first road health defence physiology mode, for example GI epithelium barrier.The consequence that the damaging dosage radiation that the repeat administration of lactoferrin compositions can cause health to absorb causes weakens.

[0073] in certain embodiments, the imagination immune system, no matter be local, system or mucosa, all strengthen by lactoferrin compositions stimulating cytokine and/or chemotactic factor.The typical cells factor comprises interleukin-18 and the GM-CSF in the gastrointestinal tract that known enhance immunity cell or immune stimulatory cell produce.For example, interleukin-18 strengthens natural killer cell or the T cell of the antibacterial that can kill infected wound.In specific embodiments, interleukin-18 (IL-18) strengthens CD4+, CD8+ and CD3+ cell.It is known to those skilled in the art that IL-18 be stimulate lymphocyte IFN-γ produce in interleukin 12 and the synergistic Th1 cytokine of interleukin II.Other cytokines or chemotactic factor also can be enhanced, such as but not limited to IL-12, IL-1b, MIP-3, MIP-1 or IFN-γ.Other cytokines or enzyme can be suppressed, such as but not limited to IL-2, IL-4, IL-5, IL-10, TNF-α or matrix metalloproteinase.

[0074] after the damaging radiation of absorbed dose, the damage of immunity and hemopoietic system makes the experimenter easily suffer from opportunistic infection and disease.The total leukocyte counting is the index of immune system injury traditionally.When being subjected to that all PBMC (peripheral blood lymphocytes) absolute quantity descends after the radiation, some change faster than other, cause the various hemocyte groups' of its initial proportion ratio change relatively.Therefore imagining lactoferrin compositions of the present invention can strengthen or increase PBMC, or reduces the decay of PBMC.More specifically, the change that immunocyte was formed relatively during known described damage caused circulating increases as the CD4+T lymphocyte, and bone-marrow-derived lymphocyte reduces, and the remarkable increase of natural killer cell.This variation causes the immunomodulating obstacle, and suppresses the immune responsiveness that antigen is attacked.Therefore, lactoferrin compositions of the present invention can be corrected or actively be changed the immune disorder that generation is replied in radiation damage.

[0075] in further embodiment, cytokine, as interleukin-18 or granulocyte/M-CSF, generation or activity that can the immune stimulatory cell.Described immunocyte includes but not limited to T lymphocyte, natural killer cell, macrophage, dendritic cell and polymorphonuclear cell.More particularly, described polymorphonuclear cell is a neutrophilic granulocyte, and described T lymphocyte is selected from CD4+, CD8+ and CD3+T.

[0076] further, imagination lactoferrin compositions cell cultured supernatant produces MIP-3 α.The known system of defense that helps health by its anti-microbial property of lactoferrin.In addition, evidence suggests that restructuring lactoferrin (rhLF) causes congenital widely sample immunoreation when oral.Innate immune system is health anti-adverse environment " the first line of defence ", comprises various effectors and cell mechanism.This innate immune response at first may be by the known enterocyte surface that is present in, as pattern recognition receptor, IL-1 receptor and the receptor " detection system " of " street cleaner " receptor regulate.There are the specific structural features of molecule in these receptor identifications and response.Consequently, signal transduction pathway may start (for example NF κ B, Wnt etc.) in the various cells, causes the comprehensive arrangement of health to the cell response of ubiquity situation biology (as infecting).Just generate important chemokine---be with regard to the MIP-3-α, RhLF and caused the hepatocellular similar response of vitro human from the component of rhLF.

[0077] in addition, when per os used, lactoferrin was also very important to the effect of the integrity of keeping the GI shielding, because this can cause antibacterial transhipment and endotoxin to cross the decay of GI epithelial cell process.Therefore, the probability of serious systemic infection development after the lactoferrin reduction radiation.In addition, lactoferrin also can reduce the total microorganism burden of intestinal, reduces the amount of free endotoxin (LF is in conjunction with endotoxin), and reduces the degree of these " unwelcome materials " migration.

V. therapeutic alliance

[0078] in order to improve the effectiveness of lactoferrin compositions of the present invention, may preferably provide protection or treatment ionizing radiation effective substances to unite use compositions of the present invention and other.These other radiation protection compositions can provide with the binding capacity of effective promotion therapeutic effect.This process may relate to uses lactoferrin compositions of the present invention and described material or a plurality of factor simultaneously.This can be by the pharmacological preparation of using single compositions or comprising two materials, or by using two different compositionss or preparation, thereby side by side or in the enough approaching time give to cause the overlapping of this effect, one of them compositions comprises lactoferrin compositions, other comprise described second material.

[0079] in addition, lactoferrin compositions of the present invention can with a few minutes to the interval in several weeks, prior to or after other radioprotective substances and/or treatment.Radioprotective substance and lactoferrin compositions use respectively or the embodiment used in, guarantee generally between each time of administration that considerable time can not lose efficacy, described like this material and lactoferrin compositions still can be brought into play favourable joint effect.Under these circumstances, imagine a kind of material and can in each other about 1-14 days, contact zone to be treated and/or be applied to experimenter to be treated, more preferably in each other about 12-24 hour.In some cases, obviously the extended treatment time limit is desirable, but several days (2,3,4,5,6 or 7) are arrived in the expiration date in several weeks (2,3,4,5,6,7 or 8) between using separately.

[0080] in specific embodiments, can be intended to reduce the treatment of damaging radiation effect with other with the lactoferrin treatment, for example with granulocyte stimulating factor (G-CSF) (Filgrastim/ (Neupogen)), or and amifostine, or share with other materials that are intended to treat the radiation damage consequence.

A. the chemical compound that contains sulfydryl

[0081] example that can be used for the sulfydryl of radioprotective substance includes but not limited to cysteine, cysteamine, cystamine, AET and 2-MEG (MEG).Mercapto-amine also is the active substance that reduces temperature and physiological pH.The dosage of all cpds reduces the factor, and (dosereduction factor, DRF) scope from 1.4 to 2.0.These compounds characteristics are sulfhydryl compound (SH) and amine (NH ₂) with 2 carbon atoms separate.

[0082] other sulfydryl free radical that can be used as radioprotective substance includes but are not limited to thiourea, thiouracil, dithiocar-bamate, dithiooxamide, thiazoline, sulfoxide and sulfone.

B. pharmacological agents

[0083] pharmacological agents that can be used as radioprotective substance can comprise anaesthetic and alcohol, analgesics (for example morphine, heroin, sodium salicylate), tranquilizer, cholinergic drug (as acetylcholine, methacholine chloride), epinephrine and norepinephrine, dopamine, histamine, serotonin, glutathion, vitamin C, vitamin E and hormone (as estrogen).

C. other material

[0084] other radioprotective substance can include but not limited to cyanide, nucleic acid derivative (as ATP), 1080 (Monsanto), p-amino-propiophenone (PAPP), melittin, endotoxin, imidazoles, 3 ', 5 '-cyclic adenosine monophosphate (cAMP), antibiotic, lipid (as olive oil), erythropoietin, carbon monoxide (with the hemoglobin competition), mercamine hydrochloride (MEA), S-(2-amino-ethyl) dibastic sodium phosphate (WR-638), S-2-(3-amino propyl amino) ethyl phosphonic acid (WR-2721), S-2-(3-amino propyl amino) propyl group phosphoric acid (WR-44923), nature polyamines putrescine (1, the 4-diaminobutane), spermidine and spermine.

[0085] other radioprotector includes but not limited to nitroxide Tempol (4-hydroxyl-2,2,6,6-tetramethyl piperidine-1-oxygen base (oxyl)), calcium antagonist (diltiazem, nifedipine and nimodipine), stobadine and bacterial endotoxin.

D. immunomodulator

[0086] immunomodulator is the another kind of radioprotector that can improve the raying animal survival.With regard to its radiation protection effect, the cytokine of broad research is: interleukin-1 (IL-1), tumor necrosis factor-alpha (TNF-α), granulocyte colony-stimulating factor (G-CSF) and granular leukocyte macrophage CSF (GM-CSF).

[0087] another is that the immunomodulator of radioprotector is the AS101 (trichlorine (dioxy ethylene-O-O ') is closed ammonium tellurate) that stimulates various cytokines to generate, and shows the radiation protection activity in mice.

VI. embodiment

[0088] the following examples are included into, with the explanation preferred embodiment of the invention.It will be understood by those skilled in the art that disclosed technology is found for the inventor among the embodiment after the prior art, bring into play good action in practice of the present invention, therefore can be considered makes up the preference pattern of its practice.Yet those skilled in the art understand according to the disclosure, can make many changes in the disclosed specific embodiments, and still obtain alike or similar result and unswerving the spirit and scope of the present invention.

Embodiment 1

Oral lactoferrin is to the effect of radiation-induced mouse death rate

[0089] mice (n=10) is subjected to the ionizing radiation of about 10Gy whole body fatal dose.Mice is immediately with dosage 2.9g/m after the radiation ²Lactoferrin or solvent placebo irritate stomach and handle.After the radiation every mice of this dosage once a day, totally 30 days.In the time of the 30th day, placebo group has 4 survival mice, 7 survival mice of TLF processed group, and promptly because TLF treats, survival increases by 75% (Fig. 1) relatively.In addition, the mice that TLF handles in the whole research shows better clinical indication.

Embodiment 2

Oral lactoferrin to the radiation of sustaining damage property after in the mice circulation leukocyte restore Effect

[0090] mice (n=10) is subjected to the ionizing radiation of about 5Gy whole body non-lethal dose.Mice is immediately with dosage 2.9g/m after the radiation ²Lactoferrin or solvent placebo irritate stomach and handle.After being subjected to radiation every mice of this dosage once a day, totally 34 days.Not radiation, untreated mice are with comparing.Each time point before the radiation and after the radiation is with the leukocyte total amount of facs analysis mice blood sample.Find and contrast, handle except improving the mice survival, also the resume speed (see figure 2) of white blood cell count in the acceleration cycle with TLF with solvent processing.The recovery of this raising may make mice that superinfection is had higher resistance.

Embodiment 3

Oral lactoferrin is to the effect of radiation-induced mice damage

[0091] mice (n=12) is subjected to the ionizing radiation of about 5Gy whole body non-lethal dose.Mice is immediately with dosage 2.9g/m after the radiation ²Lactoferrin (n=6) or solvent placebo (n=6) irritate stomach and handle.After being subjected to radiation every mice of this dosage once a day, totally 30 days.Each interval is collected the mice blood sample, analyzes the cell of mice blood and forms.Compare with the placebo treatment mice, the mouse immune system cells quantity normalization that lactoferrin is handled gets faster (table 1).

Table 1

Embodiment 4

Oral lactoferrin to radiation after the effect of health status and mortality rate

[0092] mice (n=20) is subjected to the ionizing radiation of whole body 6Gy dosage.Mice is immediately with him lactoferrin (2.9mg/kg) or the processing of placebo filling stomach after the radiation.After the radiation every mice of this dosage once a day, totally 30 days.His lactoferrin makes the survival of raying mice increase relative 50% (when promptly research finished, the mice of TLF processed group survival (6) was the twice of placebo group (3)).In the research process, the health status of mice uses Morton method (Morton 1999) to estimate every day before administration.The single numerical value of health status is measured with following parameters.

Active 1-is normal; 2-reduces; 3-is low

1-is normal for the back of a bow; 2-is medium; 3-at utmost

It is normal to stick up mao 1-; 2-is slight; 3-is medium; 4-at utmost

It is normal to breathe 1-; 2-is painstaking; 3-is shallow; 4-is rapid

Vigilance 1-is normal; 2-reduces; 3-is low

Body weight 1-increases; 2-reduces

1-is normal in dehydration; 2-is medium; 3-at utmost

Diarrhoea 1-does not have; 2-has

Polyuria (wetting) 1-does not have; 2-has

[0093] above-mentioned condition score value scope is 9 to 26.Dead animal gives final scoring 30.The result is illustrated in Fig. 3.Use repeated measure ANOVA, the counting statistics data.After these results showed the 6Gy radiation, placebo group was significantly improved (p=0.0259) on the healthy score value statistics of his lactoferrin group relatively.

Embodiment 5

Oral and intravenous injection lactoferrin is to the dose dependent of radiation-induced dead mouse Protection

[0094] mice (n=20/ group) is subjected to the ionizing radiation of about 10Gy whole body fatal dose.Mice is immediately with dosage 0 (solvent), 0.19,0.86,2.0 and 2.9mg/m after the radiation ²Lactoferrin irritates stomach or intravenous injection is handled.Mice once-a-day administration after the radiation, totally 30 days.The assessment lactoferrin is to the mouse death rate that is subjected to the fatal dose caused by ionizing radiation and the effect of holistic health thereof.With respect to control animal, animal dead rate and/or health status that lactoferrin is handled improve.

Embodiment 6

The dose dependent oral and the intravenous injection lactoferrin damages radiation-induced mice Protection

[0095] mice (n=6/ group) is subjected to the ionizing radiation of about 5Gy whole body non-lethal dose.Mice is immediately with dosage 0 (solvent), 0.19,0.86,2.0,2.9 and 5.8mg/m after the radiation ²Lactoferrin irritates stomach or intravenous injection is handled.Be subjected to every mice once-a-day administration after the radiation, totally 30 days.Each interval is collected the mice blood sample, analyzes the cell of mice blood and forms.The assessment lactoferrin is formed the mouse death rate that is subjected to caused by ionizing radiation, its blood and the effect of holistic health.With respect to control animal, the animal dead rate that lactoferrin is handled, hemocyte recover and/or health status all improves.

Embodiment 7

Oral and the intravenous injection lactoferrin of different schemes is to the effect of radiation-induced damage

[0096] mice (n=6/ group) is subjected to the ionizing radiation of about 5Gy whole body non-lethal dose.Oral immediately or intravenous injection lactoferrin processing mice after preceding 24 hours of radiation and the radiation.In different mice groups, described animal a) a day twice, dosage 1.45mg/m ²Or 2.9mg/m ², b) once a day, 2.9mg/m ²Or 5.8mg/m ², c) every other day once, dosage 2.9mg/m ²Or 5.8mg/m ², or d) weekly, dosage 2.9mg/m ²Or 5.8mg/m ²Lactoferrin is handled.Be subjected to the radiation post processing and continue 30 days.Each interval is collected blood sample, analyzes the cell of mice blood and forms.The assessment lactoferrin is formed the mouse death rate that is subjected to caused by ionizing radiation, its blood and the effect of holistic health.With respect to control animal, mortality rate, hemocyte recovery and/or health status that lactoferrin is handled animal all improve.

Embodiment 8

Oral and the intravenous injection lactoferrin of different schemes is to the effect of radiation-induced death

[0097] mice (n=10/ group) is subjected to the ionizing radiation of about 10Gy whole body fatal dose.Oral immediately or intravenous injection lactoferrin processing mice after preceding 24 hours of radiation and the radiation.In different mice groups, described animal a) a day twice, dosage 1.45mg/m ²Or 2.9mg/m ², b) once a day, 2.9mg/m ²Or 5.8mg/m ², c) every other day once, dosage 2.9mg/m ²Or 5.8mg/m ², and d) weekly, dosage 2.9mg/m ²Or 5.8mg/m ²Lactoferrin is handled.Be subjected to the radiation post processing and continue 30 days.The lactoferrin of assessment various dose and dosage regimen is to the mouse death rate that is subjected to the fatal dose caused by ionizing radiation and the effect of holistic health thereof.With respect to control animal, mortality rate and/or the health status of the animal that lactoferrin is handled all improve.

Embodiment 9

Oral and intravenous injection lactoferrin to the sublethal dose ionizing radiation after the mice secondary The protective effect of infecting

[0098] mice (n=10/ group) is subjected to the ionizing radiation of about 5Gy whole body sublethal dose.Mice is immediately with dosage 2.9mg/m after the radiation ²Or 5.8mg/m ²Stomach irritated by lactoferrin or placebo or intravenous injection is handled.Be subjected to after the radiation every mice every day once, give 30 days to lactoferrin.After the radiation three (3) days, the mice stomach was raised dosage～10 ¹²The product enterotoxin E .coli of CFU/kg.Assessment various dose lactoferrin is to being subjected to the effect of ionizing radiation mortality of mice, infection biological body and mice holistic health.With respect to control animal, mortality rate and/or the health status of the animal that lactoferrin is handled all improve.

Embodiment 10

Oral different folders with intravenous injection source lactoferrin is to the work of radiation-induced mice damage With

[0099] mice (n=24) is subjected to the ionizing radiation of about 5Gy whole body non-lethal dose.Be subjected to mice after the radiation (every group lactoferrin 6) immediately with dosage 2.9mg/m ²Various lactoferrin compositions (people of separate sources and Bovine Lactoferrin-Agennix, Ventria, Jarrow and Pharming) or solvent placebo (n=6) is irritated stomach or intravenous injection is handled.Be subjected to every group of once-a-day administration of this dosage after the radiation, totally 30 days.Each interval is collected the mice blood sample, analyzes the cell of mice blood and forms.Assessment people of separate sources or Bovine Lactoferrin are formed mouse death rate, its blood of being subjected to the fatal dose caused by ionizing radiation and the effect of holistic health.With respect to control animal, the animal dead rate that lactoferrin is handled, hemocyte recover and/or health status all improves.

Embodiment 11

The oral lactoferrin radioprotective is induced the protective effect of beasle dog death

[0100] 10 beasle dogs are used in this research, and regardless of sex, The median age is individual month of 9 (the ranges of age 7 to 32).Five beasle dogs are subjected to TLF and non-total body radiation (TBI).The group of five beasle dogs is subjected to 400cGy TBI, and gives TLF 2 in little, and lactoferrin dosage is 0.2,0.4 or 0.8g/m ²

Isolated when [0101] Canis familiaris L. arrives, the examination disease indication, and before emitting use, observed at least 1 month.Canis familiaris L. deinsectization, and inoculation rabies, canine distemper, leptospirosis, hepatitis and parvovirus vaccine.In the standard laboratory experiment, beasle dog stays in the facility of U.S. laboratory animal certification committee (American Association for Accreditation ofLaboratory Animal Care) approval, and provide commercial dog feed and chlorination tap water, ad lib.The animal feeding district remains on 70 ± 2 ℉, and the 50%-10% relative humidity at least per hour changes 15 times 100% condition fresh air.Canis familiaris L. is in light/dark full gloss spectrum illumination period of 12 hours, does not have dusk.This research flow process is through the care of animal and use committee (Institutional Animal Care and Use Committee) approval.

[0102] all Canis familiaris L.s are accepted 10cGy/ minute 400cGy TBI from two relative 60Co sources.This sky of TBI is designated as 0 day.TBI obtains packed cell volume, reticulocyte, leukocyte, platelet and the differential counting of every day before and afterwards.With the tissue check all dead Canis familiaris L.s are carried out conventional autopsy.

[0103] every day, peripheral blood cells counting logarithm value was mapped to the time.For being subjected to radiating Canis familiaris L., calculate the logarithmic average of cytometry every day that LF-handled and contrasted Canis familiaris L..On the chart, these results with connect every group every day average cube tooth bar curve display, be expressed as the logarithmic average of cytometry of time function.By the logarithmic average of cytometry is modeled as 5 order polynomials, there is constant difference time between class mean, relatively LF handle with matched group TBI after cytometry distribute, with independent covariance matrix use Generalized estimating equation (GEE) technology, detect this constant whether with 0 there were significant differences.For at least 18 days Canis familiaris L. of survival, calculate the cytometry minimum point.Use Kruskal-Wallis to detect the difference between two groups in platelet and the neutrophilic granulocyte counting minimum point.With respect to control animal, the animal hemocyte that lactoferrin is handled recovers to improve.

Embodiment 12

The oral lactoferrin radioprotective is induced the protective effect of non-human primate death

[0104] male Rhesus Macacus Macaca mulatta average weight 4.35 ± 0.32kg stays in the independent Rotating Stainless Steel Cage in the conventional receptacle of animal facility of U.S. laboratory animal certification committee (American Association forAccreditation of Laboratory Animal Care) approval.Per hour change 10 times 100% fresh air for monkey provides, condition is 72 ° ± 2 ℉, relative humidity 50% ± 20%, and remain in light/dark full gloss spectrum illumination period of 12 hours, there is not dusk.For monkey provides the commercial primate feedstuff that has added fresh fruit and tap water, ad lib.The laboratory animal nursing that experimental evidence the National Research Council laboratory animal The Study on Resources is prepared and the principle of guide for use explanation are carried out.

[0105] pre-adaptation after date, monkey is used the 250kVpx-ray on LuciteR constraint chair, 13cGy/ minute postero-anterior position, dosage (300cGy) anteroposterior position in the Rotate 180 ° is finished the one-sided irradiation of 600cGy center line irradiated tissue altogether.Use paired 0.5cm3 ionization chamber, use correction factor to carry out dosimetry from National Institute of Standards and Technology (NationalInstitute of Standards and Technology).

[0106] use two test group, every group of 5 animals, animal were shone in the time of the 0th day and the random assortment Therapeutic Method: A) contrast (n=5) oral solvent (PBS) contrast and B) the oral 1.5mg/m of LF ², once a day, 30 days.Irradiation back monitoring complete blood count 40 days, the assessment neutrophilic granulocyte reduces the persistent period of (ANC＜500/ μ l) and thrombocytopenia (PLT＜20,000/ μ l).Behind the TBI, get peripheral blood check whole blood (Sysmex K-4500 from saphena; Long Grove, IL) and differential counting (Wright-Giemsa Stain, Ames Automated Slide Stainer; Elkhart, IN) 40 days.

[0107] all animals are subjected to the clinical support be made up of required antibiotic and liquid.Give gentamycin (Elkin Sinn, AH Robbins branch company in first 7 days treatment phases; Cherry Hill, and NJ) (10mg/ days, intramuscular injection once a day), gives BaytrilR (Bayer Corporation during the whole antibacterial therapy; Shawnee Mission, KS; Http:// www.bayerus.com) (10mg/ day, intramuscular injection, every day).The continuous administration antibiotic is kept WBC 〉=1,000/ μ l for three days on end until animal, and reaches ANC 〉=500/ μ l.Compare with control animal, the animal hemocyte that lactoferrin is handled recovers and/or health status is improved.

Embodiment 13

The oral lactoferrin radioprotective is induced the protective effect of non-human primate death

[0108] non-human primate (n=10/ group) is subjected to the ionizing radiation of about 6Gy whole body fatal dose.Primate is immediately with dosage 1.5mg/m after being subjected to radiation ²Lactoferrin or solvent placebo are irritated stomach and are handled.Be subjected to primate once-a-day administration after the radiation, totally 30 days.The assessment oral lactoferrin is to increasing the therapeutical effect of animals survived.With respect to control animal, the animal dis motility rate that lactoferrin is handled improves.

Embodiment 14

The oral lactoferrin radioprotective is induced the dosage of non-human primate (NHPS) damage The dependency protective effect

[0109] non-human primate (n=6/ group) is subjected to the ionizing radiation of about 5Gy whole body non-lethal dose.Primate is immediately with dosage 0 (solvent), 0.36,0.7 and 1.5mg/m after the radiation ²Lactoferrin is irritated stomach and is handled.Be subjected to each primate once-a-day administration of described dosage after the radiation, totally 30 days.Each interval is collected the blood sample of primate, analyzes the cell of primate blood and forms.The assessment lactoferrin is formed the NHPs mortality rate that is subjected to caused by ionizing radiation, its blood and the effect of holistic health.With respect to control animal, mortality rate, hemocyte recovery and/or health status that lactoferrin is handled animal all improve.

List of references

[0110] all patents mentioned in the description and publication all signify the present invention relates to those skilled in the art's level.All patents and publication are all incorporated by reference herein, and its degree all specifically and individually shows incorporated by reference as each independent publication.

[0111] Artym J, Zimecki M, Kruzel ML:Reconstitution of thecellular immune response by lactoferrin in cyclophosphamide-treatedmice is correlated with renewal of T-cell compartment (restructuration of lactoferrin cellular immune responses is relevant with the renewal of T cell compartment in the mice of cyclophosphamide processing) .Immunobiology, 2003; 207:187-205.

[0112] Bagby GC Jr, McCall E, Layman DL:Regulation ofcolony-stimulating activity production.Interactions of fibroblasts, mononuclear phagocytes, the and lactoferrin (adjusting that colony stimulating activity produces.The interaction of fibroblast, mononuclear phagocyte and lactoferrin) .J ClinInvest, 1983; 71:340-44

[0113] Debbabi H, Dubarry M, Rautureau M, Tome D:Bovinelactoferrin induces both mucosal and systemic immune response inmice (mucosa and the systemic immunity of Bovine Lactoferrin inducing mouse are replied) .J Dairy Res, 1998; 65:283-93

[0114] Ellison RT III, Giehl TJ, LaForce FM:Damage of theouter membrane of enteric gram-negative bacteria by lactoferrin andtransferrin.Infect Immun (lactoferrin and transferrins are to the damaging action of intestinal gram negative bacteria adventitia), 1988; 56:2774-81

[0115] Hashizume S, Kuroda K, Murakami H:Identification oflactoferrin as an essential growth factor for human lymphocytic celllines in serum-free medium (identifying lactoferrin in serum-free medium is the essential growth factor of human lymphocyte) .Biochim Biophys Acta, 1983; 763:377-82

[0116] Kyte J, Doolittle RF.A simple method for displaying thehydropathic character of a protein (straightforward procedure of the hydrotherapy feature of display protein) .J Mol Biol.1982 May 5; 157 (1): 105-32.

[0117] Lonnerdal B, Iyer S:Lactoferrin:molecular structure andbiological function (lactoferrin: molecular structure and biological function) .Annu RevNutr, 1995; 15:93-110

[0118] Mikogami T., Heyman M., Spik G.and Desjeux J.F. (1994) Apical-to-basolateral transepithelial transport of humanlactoferrin in the intestinal cell line HT-29cl.19A (human lactoferrin is to be transported by top to end transepithelial among the HT-29cl.19A at enterocyte) .Am.J.Physiol.267:G308-315

[0119] Morton DB.Refinement of in vivo tests (refinement of in vivo test) .Dev Biol Stand.1999; 101:187-93

[0120] Rado TA, Bollekens J, St Laurent G et al:Lactoferrinbiosynthesis during granulocytopoiesis (the lactoferrin biosynthesis in the granulocyte generation) .Blood, 1984; 64:1103-9

[0121] Rich IN, Sawatzki G:Lactoferrin, the signalfor colonystimulating factor production? Negative-feedback regulation versussupply and-demand regulation of myelopoiesis (lactoferrin, the signal that colony stimulating factor produces? the negative feedback of myelopoiesis is regulated supply and demand is regulated) .BloodCells, 1989; 15:400-6

[0122] Rich IN:The macrophage as a production site forhematopoietic regulator molecules:sensing and responding to normaland pathophysiological signals (macrophage is the generation site that molecule is regulated in hemopoietic: to the sensing and the response of normal and Pathophysiology signal) .Anticancer Res, 1988; 8:1015-40

[0123] Wakabayashi H, Abe S, Okutomi T et al:Cooperativeanti-Candida effects of lactoferriin or its peptides in combination withazole antifungal agents (the collaborative anti-candida albicans effect of lactoferrin or its peptide and the coupling of pyrroles's antifungal agent) .Microbiol Immunol, 1996; 40:821-25

[0124] Zimecki M, Mazurier J, Machnicki M et al:Immunostimulatory activity of lactotransferrin and maturation of CD4-CD8-murine thymocytes (maturing of the immunostimulatory activity of Lactotransferrin and CD4-CD8-Mus thymocyte cell) .Immunol Lett, 1991; 30:119-23

[0125] Zimecki M, Mazurier J, Spik G, Kapp JA:Humanlactoferrin induces phenotypic and functional changes in murinesplenic B cells (human lactoferrin is induced the phenotype and the changing function of Mus spleen B cell) .Immunology, 1995; 86:122-27

[0126] Zimecki M, Spiegel K, Wlaszczyk A et al:Lactoferrinincreases the output of neutrophil precursors and attenuates thespontaneous production of TNF-alpha and IL-6 by peripheral bloodcells (lactoferrin improves the output of neutrophilic leukocyte precursor and weakens the spontaneous generation of peripheral blood cells to TNF-α and IL-6) .Arch Immunol Ther Exp (Warsz), 1999; 47:113-18

[0127] Nuijens et al. (U.S. Patent number 6,333,311)

Though the present invention and favourable part thereof are described in detail, should be appreciated that [0128] this paper can make various changes, replacement and variation, and does not depart from as defined spirit of the present invention of additional claim and scope.In addition, the application's scope is not limited to the particular of process, machinery, manufacturing, material composition, mode, method and step in this description.As those of ordinary skills from of the present invention open understand easily, can play same function in fact with corresponding embodiment described herein or obtain process, machinery, manufacturing, material composition, mode, method or the step of identical result in fact, existing or development later on, all can be used according to the invention.Therefore, additional claim is intended to comprise such process, machinery, manufacturing, material composition, mode, method or step in its scope.

Claims (43)

1. treatment is subjected to radiating experimenter's method, and it comprises the step that the lactoferrin compositions of effective dose is applied to the experimenter, and wherein said lactoferrin compositions reduces sickness rate and/or the mortality rate that is subjected to radiating experimenter.

2. the described method of claim 1, described lactoferrin compositions is used before being subjected to radiation.

3. the described method of claim 1, described lactoferrin compositions is used after being subjected to radiation.

4. the described method of claim 1, wherein said lactoferrin compositions is dispersed in the pharmaceutically acceptable carrier.

5. the described method of claim 1, the amount of the described lactoferrin compositions of wherein using is about 0.01 to 2.0g/kg every day.

6. the described method of claim 1, the amount of the described lactoferrin compositions of wherein using is about 0.01 to 0.5g/kg every day.

7. the described method of claim 1, the oral or intravenous administration of wherein said lactoferrin compositions.

8. the described method of claim 7, wherein said lactoferrin compositions is with the liquid preparation administration.

9. the described method of claim 7, wherein said lactoferrin compositions is with the solid preparation administration.

10. the described method of claim 9, wherein said solid preparation comprises enteric coating.

11. the described method of claim 1, wherein said lactoferrin compositions topical.

12. the described method of claim 1, wherein said radiation is selected from ²³⁵U, ¹³¹I, ¹²³I, ⁹⁹Tc, ²⁰¹Th, ¹³³Xe, ¹²⁵I, ⁶⁰Co and ¹³⁷Cs, ⁶⁰Co, ¹³⁷Cs, ¹⁹²Ir, ³²P, ⁹⁰Sr, ²²⁶Ra and combination thereof.

13. treatment is subjected to the method for the sequela that the doses ionizing radiation caused, it comprises by orally give effective dose lactoferrin compositions, replenishes the step of experimenter's mucomembranous immune system.

14. strengthen the method for the gastrointestinal tract mucous immunne response of experimenter that is subjected to the absorbed dose ionizing radiation, it comprises the step of orally give effective dose lactoferrin compositions.

15. the described method of claim 14, the generation of wherein said lactoferrin compositions stimulating cytokine or chemotactic factor.

16. the described method of claim 14, wherein said lactoferrin compositions causes the inhibition of cytokine or chemotactic factor.

17. the described method of claim 15, wherein said cytokine are selected from interleukin-18 (IL-18), interleukin 12 (IL-12), granulocyte/M-CSF (GM-CSF) and IFN-(IFN-γ).

18. the described method of claim 15, wherein said chemotactic factor are macrophage inflammatory protein 3 α (MIP-3 α), macrophage inflammatory protein 1 α (MIP-1 α), macrophage inflammatory protein 1 β (MIP-1 β).

19. the described method of claim 16, wherein said cytokine are selected from interleukin II (IL-2), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 10 (IL-10), and tumor necrosis factor-alpha (TNF-α).

20. the described method of claim 33, wherein said lactoferrin compositions suppresses the generation of matrix metalloproteinase (MMPs).

21. the described method of claim 17, the wherein generation or the activity of interleukin-18 or granulocyte/M-CSF immune stimulatory cell.

22. the described method of claim 21, wherein said immunocyte is selected from T lymphocyte, natural killer cell, macrophage, dendritic cell and polymorphonuclear cell.

23. the described method of claim 22, wherein said polymorphonuclear cell is a neutrophil.

24. the described method of claim 22, wherein said T lymphocyte is selected from CD4+, CD8+ and CD3+T cell.

25. reduce the method for the experimenter's mortality rate that is subjected to the absorbed dose ionizing radiation, it comprises the described experimenter of lactoferrin compositions orally give with effective dose, to alleviate the step of described absorbed dose effect.

26. alleviate the method for absorbed dose radiation to experimenter's damaging action, it comprises the lactoferrin compositions of the described experimenter's effective dose of orally give, with the step of the damaging action that alleviates described absorbed dose.

27. the described method of claim 26 wherein alleviates described damage and causes described experimenter's mortality rate to reduce.

28. the described method of claim 26 wherein alleviates described damage and causes intestinal related system antibacterial, virus or fungal infection to reduce.

29. the described method of claim 26 wherein alleviates described damage and causes described experimenter's mortality rate to reduce.

30. alleviate the method for absorbed dose radiation to experimenter's damaging action, it comprises orally give described experimenter's effective dose lactoferrin compositions and radioprotective substance, with the step of the damaging action that alleviates described absorbed dose.

31. the described method of claim 30, wherein said radioprotective substance are granulocyte stimulating factor (G-CSF) (filgrastim/(Neupogen)) or amifostine.

32. treatment is subjected to the method for the sequela that the doses ionizing radiation causes, it comprises the lactoferrin compositions by the local application effective dose, replenishes the step of experimenter's mucomembranous immune system.

Be subjected to the method that the absorbed dose ionizing radiation causes experimenter's skin histology immunne response of radiation dermantitis 33. strengthen, it comprises the step of local application effective dose lactoferrin compositions.

34. the described method of claim 33, the generation of wherein said lactoferrin compositions stimulating cytokine or chemotactic factor.

35. the described method of claim 33, wherein said lactoferrin compositions causes the inhibition of cytokine or chemotactic factor.

36. the described method of claim 35, wherein said cytokine are selected from interleukin-18 (IL-18), interleukin 12 (IL-12), granulocyte/M-CSF (GM-CSF) and IFN-(IFN-γ).

37. the described method of claim 35, wherein said chemotactic factor are macrophage inflammatory protein 3 α (MIP-3 α), macrophage inflammatory protein 1 α (MIP-1 α), macrophage inflammatory protein 1 β (MIP-1 β).

38. the described method of claim 35, wherein said cytokine are selected from interleukin II (IL-2), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 10 (IL-10), and tumor necrosis factor-alpha (TNF-α).

39. the described method of claim 33, wherein said lactoferrin compositions suppress matrix metalloproteinase (MMPs) and produce.

40. the described method of claim 36, the wherein generation or the activity of interleukin-18 or granulocyte/M-CSF immune stimulatory cell.

41. the described method of claim 40, wherein said immunocyte is selected from T lymphocyte, natural killer cell, macrophage, dendritic cell and polymorphonuclear cell.

42. the described method of claim 41, wherein said polymorphonuclear cell are neutrophil.

43. the described method of claim 41, wherein said T lymphocyte is selected from CD4+, CD8+ and CD3+T cell.

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