Tocotrienol ether derivative as well as preparation method and application thereof

Abstract

The invention provides a tocotrienol ether derivative as shown in a formula I, and an isomer, a prodrug, a pharmaceutically acceptable salt, a hydrate or a solvent compound of the tocotrienol ether derivative, r is selected from hydrogen, a substituted or unsubstituted alkyl group with 1 to 10 carbon atoms, a substituted or unsubstituted alkenyl group with 2 to 10 carbon atoms, a substituted or unsubstituted alkynyl group with 2 to 10 carbon atoms, a substituted or unsubstituted aryl group with 6 to 20 carbon atoms, or a substituted or unsubstituted heteroaryl group with 5 to 12 members, a substituted or unsubstituted cycloalkyl group with 3 to 8 carbon atoms or a heterocycloalkyl group with 3 to 8 carbon atoms. The derivative has a better effect on the whole, and is expected to actually become a potential and lasting radiation protective agent aiming at H-ARS.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to a tocotrienol ether derivative, a preparation method thereof, and an application thereof. Background Art

[0002] People may unfortunately be exposed to ionizing radiation, which poses a great risk of harm to emergency responders and civilians. Among them, the hematopoietic system is highly sensitive to radiation. Therefore, the damage to the bone marrow caused by radiation will lead to hematopoietic acute radiation syndrome (H-ARS), the main feature of which is pancytopenia. If not treated in time, H-ARS victims may die within just a few weeks from sequelae such as bleeding, opportunistic infections, and anemia.

[0003] For large-scale radiation exposure incidents, an ideal radiation countermeasure should be effective, safe, stable, and easy to manage. To this end, a variety of drug strategies have been developed, aiming to develop new radiation countermeasures to protect or mitigate the harm caused by ionizing radiation. Among these drug strategies, developing a radiation countermeasure for H-ARS has become one of the focuses, with the aim of stimulating hematopoietic recovery. Different from the past approach that focused on free radical scavengers to minimize initial damage, current efforts are more concentrated on developing immunomodulators that can induce the production of endogenous G-CSF to achieve the protection and reconstruction of the hematopoietic system.

[0004] Although a special class of immunomodulators has been extensively studied and relatively well-developed due to their good radiation protection efficacy and low toxicity, such as vitamin E, their short plasma elimination half-life and low bioavailability limit their exposure in the body circulation. So far, most research work has adopted pharmaceutical formulation methods to improve the bioavailability of tocotrienols (a form of vitamin E). Vitamin E nanoformulations have shown some potential in improving their efficacy and bioavailability, but their clinical application still faces some limitations and challenges.

[0005] Therefore, it is necessary to further develop a new chemical molecule to improve its therapeutic effect and ensure its bioavailability. Summary of the Invention

[0006] The object of the present invention is to propose a tocotrienol ether derivative represented by formula I, its isomers, its prodrugs, its pharmaceutically acceptable salts, its hydrates, or its solvate compounds in view of the technical defects existing in the prior art. The formula I is as follows.

[0007]

[0008] Among them, R is selected from hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C6-C20 aryl, or substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl or heterocycloalkyl.

[0009] The said substitution means that one or more (preferably 1, 2, 3 or 4) hydrogen atoms on the group are each independently substituted by substituents selected from the following group: halogen, -CN, OR1, nitro, NR1R2, C1-C6 alkyl, halogenated C1-C4 alkyl (such as -CF3), C3-C8 cycloalkyl or heterocycloalkyl, C1-C6 alkylthio, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, C6-C12 aryl, 5-12 membered heteroaryl, COR3.

[0010] Furthermore, R is selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C6-C10 aryl, or substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted C3-C6 cycloalkyl or heterocycloalkyl.

[0011] Furthermore, the said substitution means that one or more (preferably 1, 2, 3 or 4) hydrogen atoms on the group are each independently substituted by substituents selected from the following group: halogen, -CN, OR1, nitro, NR1R2, C1-C5 alkyl, halogenated C1-C5 alkyl (such as -CF3), C3-C6 cycloalkyl or heterocycloalkyl, C1-C5 alkylthio, halogenated C1-C5 alkoxy, halogenated C1-C5 alkylthio, C6-C10 aryl, 5-12 membered heteroaryl, COR3.

[0012] Furthermore, R1 and R2 are each independently selected from H, C1-C4 alkyl; R3 is selected from NR1R2, OH, OR1.

[0013] Furthermore, R is selected from substituted C1-C6 alkyl, preferably substituted C1-C5 alkyl; wherein, the said substitution means that one hydrogen atom on the group is substituted by OH.

[0014] Specifically, the said halogen substitution is by fluorine, chlorine, bromine or iodine.

[0015] Furthermore, R is selected from:

[0016]

[0017] The present invention also provides a composition, comprising any one of the foregoing tocotrienol ether derivatives, its isomers, its prodrugs, its pharmaceutically acceptable salts, its hydrates or its solvate compounds, and a pharmaceutical carrier or excipient.

[0018] Specifically, the composition is selected from gastrointestinal dosage forms and parenteral dosage forms. The parenteral dosage forms include injection dosage forms, skin dosage forms, respiratory dosage forms, mucosal dosage forms, and cavity dosage forms. Among them, the injection dosage form can be an injection, intravenous injection dosage form, intramuscular injection dosage form, subcutaneous injection dosage form, intradermal injection dosage form, and acupoint injection dosage form; the respiratory administration can be an inhalant, aerosol; the transdermal administration can be a locally acting - topical solution, lotion, liniment, plaster, ointment, paste, and a systemically acting - transdermal absorption patch; the mucosal administration can be eye drops, nasal drops, gargle, sublingual tablets, buccal tablets, suppositories, oral films. The gastrointestinal dosage form can be a solution, syrup, emulsion, suspension, powder, infusion, pill, tablet, capsule, and enemas, suppositories, etc. for rectal administration. Preferably, it is a subcutaneous injection dosage form.

[0019] The present invention also provides a method for preparing the tocotrienol ether derivative as described above, including:

[0020] The δ-T3H esters (6a - e) are obtained through nucleophilic substitution reaction; or, subsequently, the esters 6a - e are hydrolyzed to obtain carboxylic acid derivatives 7a - e; or, the ester groups of the compounds 6a - e are reduced to hydroxyl groups to obtain the corresponding alcohol derivatives 8a - e; or, in the presence of KOH, δ-T3H methanesulfonate (9) reacts with various substituted ethanolamines to generate the target amine compounds 10a - c.

[0021] The present invention also provides a use of the tocotrienol ether derivative, its isomer, its prodrug, its pharmaceutically acceptable salt, its hydrate or its solvate as described above for (1) preparing anti-radiation drugs; (2) preparing drugs for inducing G-CSF production; (3) preparing drugs for peripheral blood recovery; (4) preparing drugs for restoring hematopoietic stem cells and progenitor cells in the bone marrow; and / or (5) preparing drugs for promoting extramedullary hematopoiesis.

[0022] The present invention also provides an active ingredient combination, and the active ingredient combination includes the following components:

[0023] (1) The tocotrienol ether derivative, its isomer, its prodrug, its pharmaceutically acceptable salt, its hydrate or its solvate as described in claim 1; and

[0024] (2) Anti-radiation drugs, such as Neupogen, romiplostim, etc.

[0025] The beneficial effects of the present invention include:

[0026] Based on the screening of δ-T3H, which shows the most potential in G-CSF induction, as a foundation, a series of δ-T3H ester derivatives were designed and synthesized through the esterification of the C-6 phenolic group. It was found that some ester derivatives of δ-T3H have a higher induction effect on G-CSF, possess strong radiation protection for lethally irradiated mice, and exhibit better pharmacokinetic and pharmacodynamic characteristics. When administered subcutaneously, the bioavailability is significantly improved, reaching more than 400% of that of δ-T3H at the highest, and overall showing better effects, thus having the potential to actually become a potential and persistent radiation protection agent against H-ARS. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Shows the structure of vitamin E and its effect on promoting G-CSF production. (A) Structures of α, β, γ, and δ-TOH and α, β, γ, and δ-T3H. (B) Effect of vitamin E on G-CSF production (n = 5). (C) Effect of δ-T3H concentration gradient on G-CSF production (n = 5). Data are presented as mean ± standard error of the mean (SEM), n = 5. Statistical evaluation was performed using one-way analysis of variance (ANOVA), followed by Dunnett's multiple comparison test. **P < 0.01 and ****P < 0.0001 indicate significant differences compared with the control group.

[0028] Figure 2 Quantitative analysis of G-CSF in the sera of mice for each compound. (A) Structures of δ-T3H derivatives. (B) Effect of δ-T3H derivatives on G-CSF production (n = 5). Data are presented as mean ± standard deviation.

[0029] Figure 3 δ-T3H and its derivatives increased the survival rate of lethally irradiated mice. Mice were randomly divided into different groups (n = 10), and then a single dose of vehicle (PEG400) or 100 mg / kg of the designated compound was administered 24 hours before 8.5 Gy total body irradiation (TBI). After lethal TBI, the survival status of the irradiated mice was observed for 30 days. This figure depicts the Kaplan-Meier survival curves of the designated compounds.

[0030] Figure 4It was δ-T3H and its derivative 8a that promoted the hematopoietic recovery of peripheral blood cells in mice irradiated with 6.5 Gy. Mice received subcutaneous injection treatment with vehicle or 100 mg / kg of δ-T3H or 8a 24 hours before 6.5 Gy total body irradiation (n = 8). The counts of white blood cells (WBCs) (A and B), neutrophils (C and D), platelets (E and F), and red blood cells (RBCs) (G and H) in peripheral blood were measured at the designated time points. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, no significant difference.

[0031] Figure 5 It was δ-T3H and its derivative 8a that accelerated the regeneration of HPSCs in the bone marrow of mice irradiated with 6.5 Gy. Mice received subcutaneous injection treatment with vehicle or 100 mg / kg of δ-T3H or 8a 24 hours before 6.5 Gy total body irradiation and were then analyzed on day 10 after irradiation (n = 8). (A) Total number of BM nucleated cells. (B) Gating strategy showing LK, LSK, LT-HSCs, ST-HSCs, and MPPs. (C and D) Frequencies of LK and LSK cells among BMNCs. (E and F) Absolute numbers of LK and LSK cells. (G-I) Frequencies of LT-HSC, ST-HSCs, and MPPs in the LSK population. (J-L) Absolute numbers of LT-HSC, ST-HSCs, and MPPs. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, no significant difference.

[0032] Figure 6 It was δ-T3H and its derivative 8a that promoted the extramedullary hematopoiesis of the spleen in mice irradiated with 6.5 Gy. Mice received subcutaneous injection treatment with vehicle or 100 mg / kg of δ-T3H or 8a 24 hours before 6.5 Gy total body irradiation and were then analyzed on day 10 after irradiation (n = 8). (A) Spleen index. (B) Total number of splenic nucleated cells. (C) Gating strategy showing LK, LSK, LT-HSCs, ST-HSCs, and MPPs. (D)(E) Frequencies of LK and LSK cells among BMNCs. (F and G) Absolute numbers of LK and LSK cells. (H and I) Frequencies of LT-HSC and ST-HSCs in the LSK population. (J and K) Absolute numbers of LT-HSC and ST-HSCs. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, no significant difference. Detailed implementation methods

[0033] As used herein, the terms "comprising", "including", and "containing" are used interchangeably and include not only closed definitions but also semi-closed and open definitions. In other words, the terms include "consisting of" and "consisting essentially of".

[0034] It should be understood that those of ordinary skill in the art can select substituents and substitution forms on the compounds of the present invention to produce chemically stable compounds, and the compounds can be synthesized by techniques known in the art and the methods described hereinafter. If substituted by more than one substituent group, it should be understood that these multiple groups can be on the same carbon or on different carbons, as long as a stable structure is produced.

[0035] As used herein, the term "substituted" or "substituent" means that a hydrogen atom on a group is replaced by a non-hydrogen atom group, provided that its valence requirements are met and a chemically stable compound is formed by the substitution, i.e., a compound that does not spontaneously undergo transformations such as cyclization, elimination, etc.

[0036] As used herein, the term "alkyl" refers to a straight-chain (i.e., unbranched) or branched saturated hydrocarbon group containing only carbon atoms, or a group combining straight-chain and branched chains. For example, C1-C4 alkyl refers to an alkyl group containing 1-4 carbon atoms, and representative examples include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.

[0037] The term "alkenyl" refers to a monovalent group of a branched or unbranched unsaturated hydrocarbon group, preferably having 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms. Preferred alkenyl groups include vinyl (-CH=CH2), n-propenyl (-CH2CH=CH2), isopropenyl (-C(CH3)=CH2), etc.

[0038] In the present invention, the term "halogen" refers to F, Cl, Br, or I.

[0039] In the present invention, the term "halogenated" means substituted by a halogen.

[0040] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated monocyclic, bicyclic or polycyclic (fused, bridged or spiro) ring system group. When a cycloalkyl is preceded by a carbon atom number limitation (such as C3-C8), it means that the cycloalkyl has 3-8 ring carbon atoms. In some preferred embodiments, the term "C3-C8 cycloalkyl" refers to a saturated or partially saturated monocyclic or bicyclic alkyl group having 3-8 ring carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, or similar groups. "Spirocycloalkyl" refers to a bicyclic or polycyclic group in which a single carbon atom (referred to as a spiro atom) is shared between monocyclic rings, and these may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. "Fused cycloalkyl" refers to a fully carbon bicyclic or polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, and one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. "Bridged cycloalkyl" refers to a fully carbon polycyclic group in which any two rings share two non-directly connected carbon atoms, and these may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system.

[0041] The term "alkoxy" refers to an R-O- group, where R is an alkyl group as defined above herein. When an alkoxy is preceded by a carbon atom number limitation, such as C1-C6 alkoxy, it means that the alkyl group in the alkoxy has 1-6 carbon atoms. Representative examples of alkoxy include (but are not limited to): methoxy, ethoxy, n-propoxy, isopropoxy, tert-butoxy, or similar groups.

[0042] As used herein, the term "alkylthio" refers to an R-S- group, where R is an alkyl group as defined above herein. When an alkylthio is preceded by a carbon atom number limitation, such as C1-C6 alkylthio, it means that the alkyl group in the alkylthio has 1-6 carbon atoms. Representative examples of alkylthio include (but are not limited to): methylthio, ethylthio, n-propylthio, isopropylthio, tert-butylthio, or similar groups.

[0043] As used herein, the term "haloalkoxy" refers to haloalkyl-O-, where the haloalkyl is as defined above. For example, halo C1-C4 alkoxy refers to a haloalkoxy containing 1-4 carbon atoms, and representative examples include but are not limited to, monofluoromethoxy, monofluoroethoxy, difluorobutoxy, or similar groups.

[0044] As used herein, the term "haloalkylthio" refers to haloalkyl-S-, where the haloalkyl is as defined above. For example, halo C1-C4 alkylthio refers to a haloalkylthio containing 1-4 carbon atoms, and representative examples include but are not limited to, monofluoromethylthio, monofluoroethylthio, difluorobutylthio, or similar groups.

[0045] The term "heterocycloalkyl" refers to a cyclic group that is completely saturated or partially unsaturated (including but not limited to, for example, a 4- to 7-membered monocyclic ring, a 7- to 11-membered bicyclic ring, or an 8- to 16-membered tricyclic system), in which at least one heteroatom is present in a ring having at least one carbon atom. When the number of members of the heterocycloalkyl is limited, it refers to the number of ring atoms of the heterocycloalkyl. For example, a 3- to 12-membered heterocycloalkyl refers to a heterocycloalkyl having 3 to 12 ring atoms, and each heterocyclic ring containing a heteroatom may carry one or more (such as 1, 2, 3, or 4) heteroatoms selected from nitrogen atoms, oxygen atoms, or sulfur atoms, where the nitrogen atom or sulfur atom may be oxidized, and the nitrogen atom may also be quaternized. The heterocycloalkyl may be attached to the residue of any heteroatom or carbon atom of the ring or ring system molecule. Typical monocyclic heterocycloalkyls include but are not limited to azetidinyl, pyrrolidinyl, oxetanyl, pyrazolinyl, imidazolinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, hexahydroazepinyl, 4-piperidinonyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholine sulfoxide, thiomorpholine sulfone, 1,3-dioxolanyl, and tetrahydro-1,1-dioxothiophene, etc. Polycyclic heterocycloalkyls include spirocyclic, fused-ring, and bridged-ring heterocyclic groups; the spirocyclic, fused-ring, and bridged-ring heterocyclic groups involved are optionally connected to other groups by a single bond, or further fused to other cycloalkyls, heterocycloalkyls, aryls, and heteroaryls by any two or more atoms on the ring.

[0046] The term "aryl" refers to a monocyclic or fused polycyclic (i.e., rings sharing adjacent carbon atom pairs) group of all-carbon atoms having a conjugated π-electron system, which is an aromatic cyclic hydrocarbon group. When the number of carbon atoms is limited in front of the aryl, such as C6-C12 aryl, it means that the aryl has 6 to 12 ring carbon atoms, such as phenyl and naphthyl. The aryl ring may be fused to other cyclic groups (including saturated or unsaturated rings), but does not contain heteroatoms such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent must be on a carbon atom of the ring having a conjugated π-electron system.

[0047] The term "heteroaryl" refers to an aromatic heterocyclic group having one or more (preferably 1, 2, 3 or 4) heteroatoms, which can be monocyclic (monocyclic) or polycyclic (bicyclic, tricyclic or polycyclic) fused together or covalently linked, and each heterocycle containing a heteroatom can carry one or more (such as 1, 2, 3, 4) heteroatoms each independently selected from the group consisting of oxygen, sulfur and nitrogen. When there is a yuan number limit in front of the heteroaryl, it refers to the number of ring atoms of the heteroaryl. For example, a 5-12 membered heteroaryl refers to a heteroaryl having 5-12 ring atoms. Representative examples include, but are not limited to: pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl and tetrazolyl, etc.

[0048] As used herein, when alone or as part of other substituents, the term "nitro" means -NO2.

[0049] As used herein, when alone or as part of other substituents, the term "cyano" means -CN

[0050] As used herein, when alone or as part of other substituents, the term "hydroxy" means -OH.

[0051] Isomers or hydrates, such as optical isomers or racemic compounds; pharmaceutically acceptable salts, which can be sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, hydrochlorides, bromides, iodides, acetates, propionates, octanoates, acrylates, formates, isobutyrates, heptanoates, decanoates, propiolates, oxalates, malonates, succinates, octanedioates, decanedioates, fumarates, maleates, 2-butyn-1,4-dioates, 3-cyclohexyn-2,5-dioates, benzoates, chlorobenzoates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, hippurates, β-hydroxybutyrates, glycolates, maleates, tartrates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, mandelates, glutamates, argininates, lysinates, etc., preferably hydrochlorides and phosphates.

[0052] In this specification, it should be construed that all substituents are unsubstituted unless explicitly described as "substituted" herein. The term "substituted" means that one or more hydrogen atoms on a specific group are replaced by a specific substituent. The specific substituent is the substituent described correspondingly in the foregoing or the substituent appearing in each embodiment. Unless otherwise specified, an arbitrarily substituted group can have a substituent selected from a specific group at any substitutable site of the group, and the substituents can be the same or different at each position.

[0053] The present invention provides a method for (1) preparing anti-radiation drugs; (2) preparing drugs for inducing the generation of G-CSF; (3) preparing drugs for peripheral blood restoration; (4) preparing drugs for restoring hematopoietic stem cells and progenitor cells in the bone marrow; and / or (5) preparing drugs for promoting extramedullary hematopoiesis. In the present invention, the term "prevention" refers to a method for preventing the onset of a disease and / or its attendant symptoms or protecting a subject from acquiring the disease. As used herein, "prevention" also includes delaying the onset of a disease and / or its attendant symptoms and reducing the risk of the subject contracting the disease.

[0054] In the present invention, the term "treatment" refers to any treatment of a disease in a mammal, including (but not limited to): (a) inhibiting the disease, i.e., slowing or preventing the development of clinical symptoms; and / or (b) alleviating the disease, i.e., causing the regression of clinical symptoms, and / or (c) reducing or eliminating the disease and / or its attendant symptoms.

[0055] Typically, the composition is a pharmaceutical composition, which comprises a compound of formula I as described in the present invention, or an isomer thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0056] In the composition of the present invention, the amount of the compound of formula I is a therapeutically effective amount, wherein "therapeutically effective amount" refers to an amount that produces a function or activity in a human and / or an animal and is acceptable to the human and / or the animal. Those of ordinary skill in the art should understand that the "therapeutically effective amount" may vary depending on the form of the pharmaceutical composition, the route of administration, the excipients used in the drug, the severity of the disease, and the combination with other drugs.

[0057] In the present invention, the dosage forms of the tocotrienol ether derivative pharmaceutical composition include (but not limited to) oral preparations, injections, and topical preparations.

[0058] Representative ones include (but not limited to): tablets, injections, infusions, ointments, gels, solutions, microspheres, and films. The term "pharmaceutically acceptable carrier" refers to one or more compatible solid, semi-solid, liquid, or gel fillers that are suitable for human or animal use and must have sufficient purity and sufficiently low toxicity. "Compatibility" means that the components in the pharmaceutical composition and the active ingredient of the drug are admixed with each other without significantly reducing the drug efficacy.

[0059] Examples of pharmaceutically acceptable carrier moieties include cellulose and its derivatives (such as methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), buffering agents, chelating agents, thickening agents, pH regulators, transdermal penetration enhancers, coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, bacteriostatic agents, pyrogen-free water, etc.

[0060] Typically, in addition to the active pharmaceutical ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers. For example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances, etc. In addition to these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers, and suspending agents, etc.

[0061] The pharmaceutical preparation should be matched with the administration route. The medicament of the present invention can also be used together with other co-therapeutic agents (including before, during, or after use). When using the pharmaceutical composition or preparation, a safe and effective amount of the drug is administered to the desired subject (such as a human or non-human mammal), and the safe and effective amount is usually at least about 10 micrograms per kilogram of body weight, and in most cases does not exceed about 8 milligrams per kilogram of body weight. Preferably, the dose is about 10 micrograms per kilogram of body weight to about 1 milligram per kilogram of body weight. Of course, the specific dose should also consider factors such as the administration route and the patient's health condition, which are within the scope of the skills of a skilled physician.

[0062] The mice of the present invention are C57BL / 6J male mice (6 - 8 weeks old, body weight 21 - 24 g), purchased from Huafukang (HFK) Biotechnology Co., Ltd. (Beijing, China).

[0063] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0064] Screening of Vitamin E with Radiation Protection Properties

[0065] We compared the G-CSF induction levels of α, β, γ, and δ-TOH, as well as the related tocotrienols. Serum samples were collected 24 hours after dosing or vehicle administration and analyzed using a mouse G-CSF ELISA kit. As Figure 1 shown, the doses of 100 mg / kg of α, β, γ, and δ-TOH, as well as α-T3H, did not significantly increase G-CSF induction compared to the vehicle. In contrast, β-T3H administered at a dose of 100 mg / kg led to a moderate increase in G-CSF induction, while γ and δ-T3H significantly increased G-CSF levels. Then, a dose-dependent test was performed on δ-T3H at doses of 12.5, 25, 50, 100, and 200 mg / kg. The doses of 100 and 200 mg / kg were significantly effective in inducing G-CSF. The 50 mg / kg dose had a moderate effect on δ-T3H, but the lower doses lost their effect. These results indicate that δ-T3H was found to be the most effective and worthy of further structural studies to improve radioprotective efficacy.

[0066] Preparation of δ-T3H derivatives

[0067] As shown below, the δ-T3H ester compounds (6a-e) were obtained by nucleophilic substitution reaction, and then the ester compounds 6a-e were hydrolyzed to obtain the carboxylic acid derivatives 7a-e. The ester groups of the compounds 6a-e were reduced to hydroxyl groups under the action of lithium aluminum hydride to obtain the corresponding alcohol derivatives 8a-e. The compounds 10a-c were synthesized from the common starting material δ-T3H methanesulfonate (9), which can be prepared from δ-T3H in a one-step reaction. In the presence of KOH, δ-T3H methanesulfonate (9) reacted with various substituted ethanolamines to generate the target amine compounds 10a-c. All the synthesized compounds were purified by silica gel column chromatography. Flash column chromatography was performed using silica gel of 230 - 400 mesh, and the reaction progress was monitored by silica gel thin layer chromatography (TLC) plates. The structures of the target compounds were characterized by 1H-NMR, 13C-NMR (Varian INOVA 600), and high-resolution mass spectrometry (HRMS) spectroscopy (Karlsruhe, Germany).

[0068]

[0069] Ethyl 2-(((R)-2,8-dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)acetate (6a)

[0070] Weigh δ-T3H (0.5 g, 1.26 mmol) and dissolve it in 30 ml of DMF. Add ethyl bromoacetate (0.52 g, 3.12 mmol), potassium carbonate (0.13 g, 3.72 mmol) and a catalytic amount of tetrabutylammonium iodide. After heating in an oil bath at 50 °C for 24 h, add ethyl acetate and water, and adjust the pH of the solution to 1 - 2. Extract with ethyl acetate, combine the organic layers, dry over anhydrous sodium sulfate, filter, and purify by FLAsh column to obtain 0.62 g of a transparent oil, with a yield of 95.3%. 1 H NMR (600 MHz, Chloroform-d) δ 6.58 (s, 1H), 6.45 (s, 1H), 5.16–5.06 (m, 3H), 4.52 (s, 2H), 4.26 (q, J = 7.1 Hz, 2H), 2.71 (q, J = 6.4 Hz, 2H), 2.15–2.09 (m, 5H), 2.06 (p, J = 7.0 Hz, 4H), 2.00–1.96 (m, 3H), 1.80 (dt, J = 13.8, 7.0 Hz, 1H), 1.73 (dt, J = 13.3, 6.5 Hz, 1H), 1.68 (s, 3H), 1.66–1.61 (m, 1H), 1.59 (d, J = 5.5 Hz, 8H), 1.55 (d, J = 9.4 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H), 1.26 (s, 3H). 13 C NMR (151 MHz, CDCl3) δ 169.53, 150.53, 146.94, 135.18, 134.98, 131.24, 127.35, 124.43, 124.28, 124.21, 120.99, 115.76, 112.47, 75.48, 66.46, 61.14, 39.80, 39.71, 39.69, 31.31, 26.77, 26.61, 25.67, 24.05, 22.63, 22.16, 17.67, 16.18, 15.99, 15.87, 14.18. HR-MS (ESI): m / z [M + H] + calcd for C 35 H 55 O3 482.3474, found 483.3462.

[0071] Ethyl 3-(((R)-2,8-dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)propionate (6b)

[0072] δ-T3H (0.5 g, 1.26 mmol) was weighed and dissolved in 30 ml of DMF. Ethyl 3-bromopropionate (0.58 g, 3.12 mmol), potassium carbonate (0.13 g, 3.72 mmol) and a catalytic amount of tetrabutylammonium iodide were added. After heating in an oil bath at 50 °C for 24 h, ethyl acetate and water were added, and the pH of the solution was adjusted to 1 - 2. The mixture was extracted with ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and purified by FLAsh column chromatography to obtain 0.69 g of a transparent oil, with a yield of 96.1%. 1H NMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.43 (s, 1H), 5.17–5.02 (m, 5H), 4.61 (q, J = 6.8 Hz, 2H), 4.21 (qt, J = 6.6, 3.3 Hz, 4H), 2.69 (p, J = 6.2, 5.8 Hz, 3H), 2.11 (d, J = 13.4 Hz, 8H), 2.06 (td, J = 14.2, 12.2, 4.9 Hz, 9H), 1.97 (dt, J = 10.5, 5.4 Hz, 8H), 1.79 (dt, J = 13.9, 7.0 Hz, 2H), 1.73 (q, J = 6.7 Hz, 2H), 1.68 (s, 6H), 1.63 (d, J = 8.1 Hz, 1H), 1.61–1.58 (m, 17H), 1.55 (s, 3H), 1.27 (s, 1H), 1.25 (t, J = 3.5 Hz, 10H). 13C NMR (151 MHz, CDCl3) δ 172.77, 150.16, 146.85, 135.18, 134.99, 131.26, 127.26, 124.44, 124.30, 124.23, 120.95, 116.35, 116.33, 113.13, 75.44, 73.59, 61.05, 39.72, 31.34, 26.78, 26.62, 25.69, 24.08, 24.03, 22.62, 22.17, 18.69, 17.68, 16.18, 16.01, 15.88, 14.17. HR-MS (ESI): m / z [M–H]– calcd for C18H21Cl2N6O3 439.1058, found 439.1042.

[0073] Ethyl 4-(((R)-2,8-dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)butanoate (6c)

[0074] δ-T3H (0.5 g, 1.26 mmol) was weighed and dissolved in 30 ml of DMF. Ethyl 3-bromobutyrate (0.63 g, 3.12 mmol), potassium carbonate (0.13 g, 3.72 mmol) and a catalytic amount of tetrabutylammonium iodide were added. After heating in an oil bath at 50 °C for 24 h, ethyl acetate and water were added, and the pH of the solution was adjusted to 1 - 2. The mixture was extracted with ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and purified by FLAsh column chromatography to obtain 0.71 g of a transparent oil, with a yield of 96.5%. 1H NMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.43 (s, 1H), 5.16–5.06 (m, 6H), 4.14 (q, J = 7.1 Hz, 4H), 3.91 (t, J = 6.1 Hz, 4H), 2.71 (q, J = 6.6 Hz, 4H), 2.49 (t, J = 7.4 Hz, 4H), 2.16–2.09 (m, 10H), 2.11–2.02 (m, 11H), 1.97 (dt, J = 11.5, 5.9 Hz, 9H), 1.80 (dt, J = 13.9, 7.0 Hz, 2H), 1.73 (dt, J = 13.3, 6.5 Hz, 2H), 1.68 (s, 7H), 1.66–1.50 (m, 21H), 1.28–1.23 (m, 13H). 13C NMR (151 MHz, CDCl3) δ 173.41, 151.39, 146.17, 135.13, 134.98, 131.26, 127.22, 124.43, 124.33, 124.22, 120.91, 115.50, 111.88, 75.34, 67.23, 60.37, 39.72, 39.70, 31.41, 30.92, 26.93, 26.77, 26.61, 25.71, 24.87, 24.06, 22.66, 22.19, 17.69, 16.19, 16.01, 15.89, 14.25. HR-MS (ESI): m / z [M + H]+ calcd for C33H51O4 511.3787, found 511.3781.

[0075] Ethyl 5-(((R)-2,8-dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)pentanoate (6d)

[0076] Weigh δ-T3H (0.5 g, 1.26 mmol) and dissolve it in 30 ml of DMF. Add ethyl bromovalerate (0.712 mmol), potassium carbonate (0.13 g, 3.72 mmol) and a catalytic amount of tetrabutylammonium iodide. After heating in an oil bath at 50 °C for 24 h, add ethyl acetate and water, and adjust the pH of the solution to 1-2. Extract with ethyl acetate, combine the organic layers, dry over anhydrous sodium sulfate, filter, and purify by FLAsh column to obtain a transparent oil (0.71 g, 96.5%). 1 H NMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.43 (s, 1H), 5.15–5.07 (m, 3H), 4.13 (q, J = 7.1 Hz, 2H), 3.88 (t, J = 5.8 Hz, 2H), 2.71 (q, J = 6.5 Hz, 2H), 2.37 (t, J = 6.9 Hz, 2H), 2.14 (s, 3H), 2.13–2.09 (m, 2H), 2.06 (p, J = 6.9 Hz, 4H), 1.97 (dt, J = 11.3, 5.7 Hz, 4H), 1.83–1.70 (m, 6H), 1.68 (d, J = 1.6 Hz, 3H), 1.66–1.61 (m, 1H), 1.59 (d, J = 6.0 Hz, 9H), 1.57–1.52 (m, 1H), 1.27–1.24 (m, 6H). 13 C NMR (151 MHz, CDCl3) δ 173.57, 151.51, 146.10, 135.13, 134.98, 131.27, 127.19, 124.43, 124.34, 124.22, 120.90, 115.46, 111.85, 75.32, 67.84, 60.27, 39.75, 39.72, 39.70, 34.03, 31.43, 28.89, 26.77, 26.61, 25.71, 24.06, 22.66, 22.19, 21.73, 17.69, 16.19, 16.01, 15.89, 14.26.

[0077] HR-MS (ESI): m / z [M+H] + calcd for C 34 H 55 O4 524.3866, found 525.3938.

[0078] Ethyl 2-(((R)-2,8-dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)-2-methylpropanoate (6e)

[0079] Weigh δ-T3H (0.5 g, 1.26 mmol) and dissolve it in 30 ml of DMF. Add ethyl 2-bromoisobutyrate (0.63 g, 12 mmol), potassium carbonate (0.13 g, 3.72 mmol) and a catalytic amount of tetrabutylammonium iodide. After heating in an oil bath at 50 °C for 24 h, add ethyl acetate and water, and adjust the pH of the solution to 1-2. Extract with ethyl acetate, combine the organic layers, dry over anhydrous sodium sulfate, filter, and purify by FLAsh column chromatography to obtain 0.68 g of a transparent oil, with a yield of 93.9%. 1 HNMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.43 (s, 1H), 5.16–5.06 (m, 6H), 4.14 (q, J = 7.1 Hz, 4H), 3.91 (t, J = 6.1 Hz, 4H), 2.71 (q, J = 6.6 Hz, 4H), 2.49 (t, J = 7.4 Hz, 4H), 2.16–2.09 (m, 10H), 2.11–2.02 (m, 11H), 1.97 (dt, J = 11.5, 5.9 Hz, 9H), 1.80 (dt, J = 13.9, 7.0 Hz, 2H), 1.73 (dt, J = 13.3, 6.5 Hz, 2H), 1.68 (s, 7H), 1.66–1.50 (m, 21H), 1.28–1.23 (m, 13H). 13 C NMR (151 MHz, CDCl3) δ 173.41, 151.39, 146.17, 135.13, 134.98, 131.26, 127.22, 124.43, 124.33, 124.22, 120.91, 115.50, 111.88, 75.34, 67.23, 60.37, 39.72, 39.70, 31.41, 30.92, 26.93, 26.77, 26.61, 25.71, 24.87, 24.06, 22.66, 22.19, 17.69, 16.19, 16.01, 15.89, 14.25. HR-MS (ESI): m / z [M+H] + calcd for C 33 H 50 O4 510.3709, found 511.3782.

[0080] 2-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)acetic acid (7a)

[0081] Weigh 6a (0.31 g, 0.417 mmol) into a 100 mL dry eggplant-shaped flask, add THF / H2O (20 mL:10 mL), and add lithium hydroxide (0.02 g, 0.834 mL). After reacting at room temperature for 8 h, the reaction is complete by TLC plate spotting. Adjust the pH of the solution to 1 - 2, extract with ethyl acetate, combine the organic layers, dry over anhydrous sodium sulfate, filter, and obtain 154.23 mg of a colorless oil by FLASH chromatography, with a yield of 71.4%. 1 H NMR (600 MHz, Chloroform-d) δ 6.60 (s, 1H), 6.47 (s, 1H), 5.16–5.06 (m, 2H), 4.59 (s, 4H), 2.73–2.67 (m, 3H), 2.19–2.01 (m, 17H), 1.97 (dd, J=8.5, 6.2 Hz, 8H), 1.81 (dt, J=14.0, 7.0 Hz, 2H), 1.74 (dt, J=13.3, 6.5 Hz, 2H), 1.69 (s, 6H), 1.68–1.51 (m, 26H), 1.26 (s, 6H). HR-MS (ESI): m / z [M-H] - calcd for C 29 H 41 O4 453.3005, found 453.3011.

[0082] 3-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)propanoic acid (7b)

[0083] Weigh 6b (0.36 g, 0.417 mmol) into a 100 mL dry eggplant-shaped flask, add THF / H2O (20 mL:10 mL), and add lithium hydroxide (0.02 g, 0.834 mL). After reacting at room temperature for 8 h, the reaction is complete by TLC plate spotting. Adjust the pH of the solution to 1 - 2, extract with ethyl acetate, combine the organic layers, dry over anhydrous sodium sulfate, filter, and obtain 105.26 mg of a colorless oil by FLASH chromatography, with a yield of 50.84%. 11H NMR (600 MHz, Chloroform-d) δ 6.58 (s, 1H), 6.46 (s, 2H), 5.16–5.06 (m, 9H), 4.66 (q, J = 6.8 Hz, 3H), 4.03–3.92 (m, 3H), 2.71 (q, J = 6.5 Hz, 6H), 2.13 (s, 8H), 2.10 (dd, J = 14.9, 7.1 Hz, 7H), 2.05 (q, J = 7.8 Hz, 13H), 2.00–1.93 (m, 11H), 1.87–1.76 (m, 5H), 1.74 (q, J = 6.7, 1.3 Hz, 3H), 1.68 (s, 5H), 1.66–1.52 (m, 39H), 1.43 (s, 2H), 1.26 (d, J = 1.9 Hz, 9H). 13 13C NMR (151 MHz, CDCl3) δ 177.38, 149.52, 147.24, 135.20, 134.99, 127.55, 127.53, 124.41, 124.24, 124.20, 121.15, 121.12, 116.44, 116.36, 113.30, 113.19, 75.56, 73.20, 39.87, 39.79, 39.72, 39.69, 31.22, 26.76, 26.59, 25.70, 24.07, 24.00, 22.59, 22.15, 18.49, 17.68, 16.20, 16.00, 15.88. HR-MS (ESI): m / z [M-H] - calcd for C 30 H 44 O4 468.3240, found 467.3171.

[0084] 4-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)butanoic acid (7c)

[0085] Weighed 6c (0.41 g, 0.417 mmol) into a 100 mL dry eggplant-shaped flask, added THF / H2O (20 mL:10 mL), and lithium hydroxide (0.02 g, 0.834 mL). After reacting at room temperature for 8 h, TLC showed that the reaction was complete. Adjusted the pH of the solution to 1 - 2, extracted with ethyl acetate, combined the organic layers, dried over anhydrous sodium sulfate, filtered, and obtained 178.26 mg of a colorless oil by FLASH chromatography, with a yield of 60.84%. 11H NMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.43 (s, 1H), 5.16–5.06 (m, 3H), 4.14 (q, J = 7.1 Hz, 2H), 3.91 (t, J = 6.1 Hz, 2H), 2.71 (q, J = 6.4 Hz, 2H), 2.49 (t, J = 7.4 Hz, 2H), 2.12 (d, J = 19.2 Hz, 5H), 2.06 (q, J = 6.9 Hz, 6H), 1.97 (dt, J = 11.2, 5.7 Hz, 4H), 1.80 (dt, J = 13.9, 6.9 Hz, 1H), 1.73 (dt, J = 13.4, 6.5 Hz, 1H), 1.68 (s, 3H), 1.66–1.52 (m, 11H), 1.26 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 173.41, 151.39, 146.17, 135.14, 134.99, 131.28, 127.22, 124.33, 124.22, 120.91, 115.49, 111.88, 75.34, 67.23, 60.38, 39.72, 39.70, 31.40, 30.92, 26.77, 26.60, 25.70, 24.86, 24.05, 22.65, 22.18, 17.69, 16.19, 16.01, 15.89, 14.24. HR-MS (ESI): m / z [M-H] - calcd for C 31 H 46 O4 482.3396, found 481.3323.

[0086] 5-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)penta-noic acid (7d)

[0087] Weighed 6d (0.41 g, 0.417 mmol) into a 100 mL dry eggplant-shaped flask, added THF / H2O (20 mL:10 mL), and then added lithium hydroxide (0.02 g, 0.834 mL). After reacting at room temperature for 8 h, TLC indicated that the reaction was complete. Adjusted the pH of the solution to 1-2, extracted with ethyl acetate, combined the organic layers, dried over anhydrous sodium sulfate, filtered, and obtained 185.32 mg of a colorless oil by FLASH chromatography, with a yield of 54.78%. 11H NMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.43 (s, 1H), 5.16–5.08 (m, 5H), 3.91–3.87 (m, 3H), 2.71 (q, J=6.4 Hz, 3H), 2.43 (t, J=6.8 Hz, 3H), 2.14 (s, 5H), 2.13–2.09 (m, 4H), 2.10–2.02 (m, 7H), 1.99–1.94 (m, 5H), 1.85–1.77 (m, 8H), 1.73 (dt, J=13.3, 6.5 Hz, 2H), 1.68 (s, 5H), 1.66–1.61 (m, 1H), 1.59 (s, 10H), 1.26 (s, 5H). 13 13C NMR (151 MHz, CDCl3) δ 179.22, 151.47, 146.16, 135.12, 134.97, 131.24, 127.19, 124.43, 124.35, 124.23, 120.92, 115.50, 111.93, 75.34, 67.84, 39.79, 39.72, 39.70, 33.59, 31.44, 28.76, 26.93, 26.77, 26.61, 25.68, 24.04, 22.66, 22.19, 21.48, 17.67, 16.16, 16.00, 15.88. HR-MS (ESI): m / z [M+Cl] - calcd for C 32 H 48 ClO4 531.3241, found 531.3232.

[0088] 2-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)-2-methylpropanoic acid (7e)

[0089] Weighed 6e (0.42 g, 0.418 mmol) into a 100 mL dry eggplant-shaped flask, THF / H2O (20 mL:10 mL), added lithium hydroxide (0.02 g, 0.834 mL), reacted at room temperature for 8 h. After the reaction was complete by TLC spotting, adjusted the pH of the solution to 1 - 2, extracted with ethyl acetate, combined the organic layers, dried over anhydrous sodium sulfate, filtered, and obtained 174.51 mg of a colorless oil by FLASH chromatography, with a yield of 56.23%. 11H NMR (600 MHz, Chloroform-d) δ 6.58 (s, 1H), 6.52 (s, 1H), 5.17–5.12 (m, 1H), 5.12–5.07 (m, 2H), 2.70 (q, J = 6.5 Hz, 2H), 1.81 (dt, J = 13.9, 7.0 Hz, 1H), 1.74 (dt, J = 13.3, 6.5 Hz, 1H), 1.69–1.61 (m, 4H), 1.59 (q, J = 4.7, 3.8 Hz, 11H), 1.51 (s, 8H), 1.27 (s, 4H). 13 13C NMR (151 MHz, CDCl3) δ 176.44, 148.77, 145.24, 135.29, 135.01, 131.26, 127.06, 124.41, 124.17, 122.46, 120.84, 120.02, 80.45, 75.79, 39.85, 39.71, 39.68, 31.17, 29.69, 26.76, 26.59, 25.68, 24.74, 24.07, 22.48, 22.15, 17.67, 16.11, 16.00, 15.85. HR-MS (ESI): m / z [M-H] - calcd for C 31 H 46 O4 481.3327, found 481.3322.

[0090] 2-(((S)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)ethan-1-ol (8a)

[0091] Weigh 6a (0.52 g, 1.02 mmol) and dissolve it in 20 mL of anhydrous ether. Add lithium aluminum hydride (57 mg, 2.04 mmol) at low temperature. After the addition, transfer it to room temperature and react for 8 h until the reaction is complete. Add water to quench the reaction, adjust the pH of the solution to 2 - 3, extract with anhydrous ether, combine the organic layers, filter, and purify by FLASH column to obtain 425.36 mg of colorless oil, with a yield of 97.71%. 11H NMR (600 MHz, Chloroform-d) δ 6.57 (s, 1H), 6.45 (s, 1H), 5.13 (t, J = 7.9 Hz, 1H), 5.09 (d, J = 5.2 Hz, 2H), 4.05 (t, J = 5.9 Hz, 2H), 3.85 (t, J = 5.8 Hz, 2H), 2.75–2.66 (m, 2H), 2.16–2.03 (m, 9H), 2.01 (d, J = 5.8 Hz, 2H), 2.01–1.94 (m, 6H), 1.80 (q, J = 6.8 Hz, 1H), 1.74 (dt, J = 13.3, 6.6 Hz, 1H), 1.69–1.61 (m, 5H), 1.59 (d, J = 6.6 Hz, 9H), 1.57–1.51 (m, 1H), 1.26 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 151.25, 146.36, 135.15, 134.98, 131.26, 127.29, 124.42, 124.31, 124.21, 120.98, 115.52, 112.00, 75.40, 66.88, 61.14, 39.75, 39.71, 39.69, 32.10, 31.40, 26.77, 26.61, 25.69, 24.04, 22.65, 22.18, 17.67, 16.16, 16.00, 15.88. HRMS-ESI (m / z): calcd. for C 29 H 44 O3[M + H] + 440.3290, found: 441.3364.

[0092] 3-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)propan-1-ol (8b)

[0093] Weighed 6b (0.54, 1.02 mmol) and dissolved it in 20 mL of anhydrous diethyl ether. Added lithium aluminum hydride (57 mg, 2.04 mmol) at low temperature. After the addition, the reaction mixture was transferred to room temperature and reacted for 8 h until the reaction was complete. Then, water was added to quench the reaction, and the pH of the solution was adjusted to 2 - 3. The mixture was extracted with anhydrous diethyl ether, and the organic layers were combined, filtered, and purified by FLASH column to obtain 431.62 mg of a colorless oil, with a yield of 93.14%. 11H NMR (600 MHz, Chloroform-d) δ 6.57 (s, 1H), 6.45 (s, 1H), 5.13 (t, J = 7.9 Hz, 1H), 5.09 (d, J = 5.2 Hz, 2H), 4.05 (t, J = 5.9 Hz, 2H), 3.85 (t, J = 5.8 Hz, 2H), 2.75–2.66 (m, 2H), 2.16–2.03 (m, 9H), 2.01 (d, J = 5.8 Hz, 2H), 2.01–1.94 (m, 6H), 1.80 (q, J = 6.8 Hz, 1H), 1.74 (dt, J = 13.3, 6.6 Hz, 1H), 1.69–1.61 (m, 5H), 1.59 (d, J = 6.6 Hz, 9H), 1.57–1.51 (m, 1H), 1.26 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 151.25, 146.36, 135.15, 134.98, 131.26, 127.29, 124.42, 124.31, 124.21, 120.98, 115.52, 112.00, 75.40, 66.88, 61.14, 39.75, 39.71, 39.69, 32.10, 31.40, 26.77, 26.61, 25.69, 24.04, 22.65, 22.18, 17.67, 16.16, 16.00, 15.88. HRMS-ESI (m / z): calcd. for C 30 H 46 O3[M + H] + 454.3447, found: 455.3522.

[0094] 4-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)butan-1-ol (8c)

[0095] 6b (0.641.02 mmol) was weighed and dissolved in 20 mL of anhydrous ether. Lithium aluminum hydride (57 mg, 2.04 mmol) was added at low temperature. After the addition, the reaction mixture was transferred to room temperature and reacted for 8 h until the reaction was complete. Water was added to quench the reaction, and the pH of the solution was adjusted to 2 - 3. The mixture was extracted with anhydrous ether, and the organic layers were combined, filtered, and purified by FLASH column to obtain 433.92 mg of a colorless oil, with a yield of 90.83%. 11H NMR (600 MHz, Chloroform-d) δ 6.58 (s, 1H), 6.46 (s, 1H), 5.16–5.08 (m, 4H), 3.99 (t, J = 5.8 Hz, 4H), 2.70 (t, J = 5.8 Hz, 8H), 2.34 (s, 13H), 1.97 (dt, J = 11.1, 5.7 Hz, 9H), 1.80 (dt, J = 13.9, 7.0 Hz, 2H), 1.73 (dt, J = 13.4, 6.5 Hz, 3H), 1.68 (d, J = 1.6 Hz, 7H), 1.66–1.61 (m, 2H), 1.59 (d, J = 5.7 Hz, 18H), 1.57–1.46 (m, 2H), 1.26 (s, 7H). 13 13C NMR (151 MHz, CDCl3) δ 151.35, 146.21, 135.14, 134.99, 131.27, 127.17, 124.43, 124.33, 124.22, 120.88, 115.59, 111.99, 75.34, 66.42, 58.45, 45.84, 39.76, 39.72, 39.70, 31.40, 26.76, 26.60, 25.70, 24.06, 22.65, 22.18, 17.69, 16.20, 16.01, 15.88. HRMS-ESI (m / z): calcd. for C 30 H 46 O3[M + H] + 468.3603, found: 469.3677.

[0096] 5-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)pentan-1-ol (8d)

[0097] Weighed 6d (0.56, 0.02 mmol) and dissolved it in 20 mL of anhydrous ether. Added lithium aluminum hydride (57 mg, 2.04 mmol) at low temperature. After the addition, the reaction mixture was transferred to room temperature and reacted for 8 h until the reaction was complete. Then, water was added to quench the reaction, and the pH of the solution was adjusted to 2 - 3. The mixture was extracted with anhydrous ether, and the organic layers were combined, filtered, and purified by FLASH column to obtain 451.87 mg of a colorless oil, with a yield of 91.84%. 11H NMR (600 MHz, Chloroform-d) δ 6.55 (s, 1H), 6.44 (s, 1H), 5.16–5.07 (m, 3H), 3.88 (t, J = 6.5 Hz, 2H), 3.67 (t, J = 6.5 Hz, 2H), 2.71 (q, J = 6.3 Hz, 2H), 2.16–2.02 (m, 9H), 1.97 (dt, J = 11.5, 5.8 Hz, 4H), 1.84–1.70 (m, 4H), 1.68 (s, 3H), 1.66–1.49 (m, 16H), 1.26 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 151.59, 146.08, 135.13, 134.98, 131.26, 127.17, 124.43, 124.35, 124.22, 120.91, 115.48, 111.90, 75.32, 68.30, 62.88, 39.76, 39.72, 39.70, 32.49, 31.44, 29.23, 26.77, 26.61, 25.69, 24.06, 22.66, 22.40, 22.19, 17.68, 16.18, 16.00, 15.88. HRMS-ESI (m / z): calcd. for C 31 H 48 O3 [M + H] + 482.3760, found: 483.3833.

[0098] 1-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)-2-methylpropan-2-ol (8e)

[0099] Weigh 6e (0.460.418 mmol) and dissolve it in 20 mL of anhydrous ether. Add lithium aluminum hydride (57 mg, 2.04 mmol) at low temperature. After the addition, transfer the reaction mixture to room temperature and react for 8 h until the reaction is complete. Quench the reaction with water and adjust the pH of the solution to 2 - 3. Extract with anhydrous ether, combine the organic layers, filter, and purify by FLASH column to obtain 410.39 mg of colorless oil, with a yield of 85.90%. 11H NMR (600 MHz, Chloroform-d) δ 6.62 (s, 1H), 6.54 (s, 1H), 5.19–5.09 (m, 3H), 3.57 (s, 2H), 2.73 (q, J = 6.4 Hz, 2H), 2.16 (s, 3H), 2.14 (s, 1H), 2.09 (p, J = 7.0 Hz, 5H), 2.00 (q, J = 7.2 Hz, 4H), 1.83 (dt, J = 13.9, 7.0 Hz, 1H), 1.77 (dt, J = 13.3, 6.5 Hz, 1H), 1.70 (s, 3H), 1.69–1.64 (m, 1H), 1.63–1.56 (m, 10H), 1.30 (s, 3H), 1.25 (s, 6H). 13 13C NMR (151 MHz, CDCl3) δ 148.43, 146.01, 135.23, 134.99, 131.26, 126.59, 124.42, 124.26, 124.20, 124.14, 121.74, 120.54, 79.74, 75.60, 70.14, 39.82, 39.72, 39.69, 31.30, 26.93, 26.76, 26.59, 25.70, 24.11, 23.18, 22.48, 22.17, 17.68, 16.12, 16.01, 15.85. HRMS-ESI (m / z): calcd. for C 31 H 48 O3[M + H] + 468.3603, found: 469.3668.

[0100] Methylsulfonyl δ-tocotrienol (9)

[0101] To δ-tocotrienol (4.0 g, 10.09 mmol) in 10 mL of dry DCM solution was added methanesulfonyl chloride (2.31 g, 20.18 mmol), pyridine (1.59 g, 20.18 mmol) and a catalytic amount of DMAP. The reaction mixture was stirred overnight at room temperature. The solvent was evaporated to dryness in vacuo. The residue was dissolved in 25 mL of ethyl acetate and washed twice with 2 * 20 mL of copper sulfate solution to remove any excess pyridine. The organic layer was dried over anhydrous sodium sulfate, the solvent was evaporated, and the sample was purified by column chromatography, eluting with 2 - 3% (v / v) ethyl acetate in hexane solution to give 4.14 g (yield 85.68%, 10% ethyl acetate in hexane) of methylsulfonyl δ-tocotrienol. 11H NMR (600 MHz, Chloroform-d) δ 6.86 (s, 1H), 6.84 (s, 1H), 5.15–5.07 (m, 3H), 3.09 (s, 3H), 2.76 (q, J = 6.5 Hz, 2H), 2.16 (s, 3H), 2.15–2.09 (m, 2H), 2.06 (p, J = 8.9, 8.0 Hz, 4H), 1.85–1.80 (m, 1H), 1.76 (dt, J = 13.4, 6.4 Hz, 1H), 1.69–1.62 (m, 4H), 1.62–1.54 (m, 10H), 1.28 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 151.05, 135.41, 135.04, 131.29, 128.03, 124.40, 124.15, 124.01, 121.54, 121.50, 119.85, 76.34, 39.89, 39.72, 39.68, 36.93, 30.83, 26.76, 26.58, 24.10, 22.41, 22.12, 17.69, 16.16, 16.01, 15.91. HRMS-ESI (m / z): calcd. for C 28 H 42 O4S [M+H] + 475.2882, found: 475.2886.

[0102] 2-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)ethan-1-amine (10a).

[0103] A mixture of ethanolamine (1.05 mmol) and potassium hydroxide (15.75 mmol) was placed in a two-necked round-bottom flask equipped with a Dean-Stark apparatus and refluxed in 10 mL of toluene for 2 h to remove water from the azeotropic mixture. Methylsulfonated δ-tocotrienol (0.5 g, 1.05 mmol) was added to the reaction mixture and refluxed at 80 °C for 48 h. The reaction mixture was taken up in ethyl acetate (100 mL), washed with water (2 × 100 mL), dried over anhydrous magnesium sulfate, and filtered. Ethyl acetate was removed from the filtrate on a rotary evaporator. The resulting residue was purified by column chromatography on silica gel (60-120 mesh) and eluted with 1-2% methanol (v / v) in chloroform to give 410.32 mg of a brown oil in 88.78% yield. 11H NMR (600 MHz, Chloroform-d) δ 6.47 (s, 0H), 6.38 (s, 0H), 5.11 (dt, J = 24.9, 8.7 Hz, 1H), 2.72–2.66 (m, 1H), 2.22–2.08 (m, 3H), 2.05 (q, J = 7.2 Hz, 2H), 2.00–1.92 (m, 2H), 1.80 (dt, J = 13.9, 7.0 Hz, 1H), 1.73 (dt, J = 13.5, 6.7 Hz, 1H), 1.68 (s, 2H), 1.63 (t, J = 8.3 Hz, 0H), 1.61–1.51 (m, 5H), 1.26 (s, 2H). 13 13C NMR (151 MHz, CDCl3) δ 147.72, 146.04, 135.16, 135.00, 131.29, 127.40, 124.43, 124.31, 124.21, 123.64, 121.26, 115.64, 112.59, 75.34, 39.72, 39.69, 31.36, 29.71, 26.76, 26.60, 25.70, 24.04, 22.48, 22.17, 17.69, 16.06, 16.01, 15.88, 14.39. HRMS-ESI (m / z): calcd. for C 29 H 45 NO2[M + H] + 440.3529, found: 440.3525.

[0104] 2-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)-N-methylethan-1-amine (10b)

[0105] A mixture of 1-methylethanolamine (1.05 mmol) and potassium hydroxide (15.75 mmol) was placed in a two-necked round-bottom flask equipped with a Dean-Stark apparatus and refluxed in 10 mL of toluene for 2 h to remove water from the azeotrope. Methylsulfonated δ-tocotrienol (0.5 g, 1.05 mmol) was added to the reaction mixture and refluxed at 80 °C for 48 h. The reaction mixture was taken up in ethyl acetate (100 mL), washed with water (2 × 100 mL), dried over anhydrous magnesium sulfate, and filtered. Ethyl acetate was removed from the filtrate on a rotary evaporator. The resulting residue was purified by column chromatography on silica gel of 60–120 mesh size and eluted with 1–2% methanol (v / v) in chloroform to give 230.43 mg of a brown oil, with a yield of 67.61%. 11H NMR (600 MHz, Chloroform-d) δ 6.58 (s, 1H), 5.09 (q, J = 5.5 Hz, 2H), 4.11–4.01 (m, 4H), 3.06 (t, J = 5.1 Hz, 4H), 2.77–2.65 (m, 4H), 2.59 (s, 6H), 2.14 (s, 7H), 2.06 (p, J = 6.9 Hz, 11H), 1.97 (td, J = 8.0, 7.0, 4.6 Hz, 11H), 1.80 (dt, J = 13.9, 7.1 Hz, 2H), 1.73 (dt, J = 13.4, 6.5 Hz, 2H), 1.68 (d, J = 1.5 Hz, 7H), 1.66–1.61 (m, 2H), 1.61–1.52 (m, 19H), 1.25 (s, 8H). 13 13C NMR (151 MHz, CDCl3) δ 150.84, 146.54, 135.18, 134.99, 131.27, 127.34, 124.41, 124.27, 124.20, 121.01, 115.60, 112.10, 75.42, 65.99, 49.92, 39.77, 39.69, 35.04, 31.35, 29.70, 26.76, 26.60, 25.70, 24.00, 22.62, 22.16, 17.68, 16.18, 16.00, 15.89. HRMS-ESI (m / z): calcd. for C 30 H 47 NO2[M + H] + 453.3607, found: 454.3679.

[0106] 2-(((R)-2,8-Dimethyl-2-((3E,7E)-4,8,12-trimethyltrideca-3,7,11-trien-1-yl)chroman-6-yl)oxy)-N,N-dimethylethan-1-amine (10c)

[0107] A mixture of N,N-dimethylethanolamine (1.05 mmol) and potassium hydroxide (15.75 mmol) was weighed and placed in a two-necked round-bottom flask equipped with a Dean-Stark apparatus. It was refluxed in 10 mL of toluene for 2 hours to remove water in the azeotropic mixture. Methylsulfonated δ-tocotrienol (0.5 g, 1.05 mmol) was added to the reaction mixture and refluxed at 80 °C for 48 hours. The reaction mixture was poured into ethyl acetate (100 mL), washed with water (2×100 mL), dried over anhydrous magnesium sulfate, and filtered. Ethyl acetate was removed from the filtrate on a rotary evaporator. The resulting residue was purified by column chromatography using silica gel of 60-120 mesh size and eluted with 1-2% methanol (v / v) in chloroform to obtain 368.95 mg of a brown oil with a yield of 80.42%. 1 H NMR (600 MHz, Chloroform-d) δ 6.58 (s, 1H), 6.46 (s, 1H), 5.16–5.08 (m, 4H), 3.99 (t, J = 5.8 Hz, 4H), 2.70 (t, J = 5.8 Hz, 8H), 2.34 (s, 13H), 1.97 (dt, J = 11.1, 5.7 Hz, 9H), 1.80 (dt, J = 13.9, 7.0 Hz, 2H), 1.73 (dt, J = 13.4, 6.5 Hz, 3H), 1.68 (d, J = 1.6 Hz, 7H), 1.66–1.61 (m, 2H), 1.59 (d, J = 5.7 Hz, 18H), 1.57–1.46 (m, 2H), 1.26 (s, 7H). 13 C NMR (151 MHz, CDCl3) δ 151.35, 146.21, 135.14, 134.99, 131.27, 127.17, 124.43, 124.33, 124.22, 120.88, 115.59, 111.99, 75.34, 66.42, 58.45, 45.84, 39.76, 39.72, 39.70, 31.40, 26.76, 26.60, 25.70, 24.06, 22.65, 22.18, 17.69, 16.20, 16.01, 15.88. HRMS-ESI (m / z): calcd. for C 31 H 49 NO2 [M+H] + 467.3673, found: 468.3836.

[0108] G-CSF induction experiment

[0109] A series of δ-T3H ether derivatives were designed and synthesized in this invention, and their effects on promoting G-CSF production were detected by ELISA test for 24 h.

[0110] Male C57BL / 6J mice were randomly divided into equal groups (n = 5) and given PEG400 (vehicle) or the derivative subcutaneously at a dose of 100 mg / kg. Twenty-four hours after dosing, the mice were anesthetized by intraperitoneal injection of sodium pentobarbital, and cardiac blood was withdrawn using a 1-ml medical syringe. Serum samples were prepared for measurement after blood coagulation and centrifugation. Using according to the manufacturer's instructions Mouse G-CSF ELISA kit.

[0111] As Figure 2 shown, after transferring the terminal carboxyl group to the hydroxyl group, compared with 7a-e, compounds 8a-e all showed a more excellent effect of inducing G-CSF production, and even higher than other types of ether derivatives, which suggests that the terminal hydroxyl group plays an important role in ensuring the activity of δ-T3H ether derivatives, and compound 8a has the highest activity.

[0112] Survival rate of δ-T3H and its ether derivatives after lethal irradiation

[0113] Inspired by the significant induction of G-CSF by δ-T3H and its derivatives in the aforementioned experiments, survival experiments were further carried out to study the radiation protection characteristics of irradiated mice. Some derivatives with good G-CSF induction activity were selected, such as 8a and 8b. After randomly dividing into groups (n = 10), mice were given subcutaneous injection of PEG400 (vehicle) or 100 mg / kg of δ-T3H and its derivatives; 24 hours after injection, the mice received 8.5 Gy total body irradiation (TBI). This radiation dose has been shown to cause severe bone marrow suppression and is associated with high mortality.

[0114] As Figure 3 shown, in the experimental group of this study, only 10 - 20% of the mice receiving 8.5 Gy total body irradiation survived 20 days after irradiation, and δ-T3H ether derivatives 8a and 8b produced 100% and 80% survival advantages respectively in the mice irradiated with 8.5 Gy.

[0115] Pharmacokinetic characteristics of δ-T3H and 8a

[0116] To verify the improvement of the absorption, distribution, metabolism and excretion (ADME) of δ-T3H ester derivatives, the pharmacokinetic characteristics of δ-T3H and its derivative (taking 8a as an example) were studied in ICR mice.

[0117] Male ICR mice were randomly divided into two groups before the experiment and fasted for 12 hours before the experiment. Then, the mice were given δ-T3H and 8a at a dose of 100 mg / kg by intravenous injection. Blood samples were collected before and after drug administration at 0.05, 0.17, 0.25, 0.75, 2, 4, 8, 24 hours (intravenous injection) and 0.25, 0.5, 1, 2, 4, 8, 24 hours (subcutaneous injection) (n=3). δ-T3H and 8a were dissolved in 5% Tween 80 and 95% aqueous solution (volume ratio). Blood samples were collected at each time point, placed in a test tube containing EDTA, and incubated at 4°C at 1.2×10 3 Centrifuge at rpm for 2 minutes to separate plasma. Take 50 μL of plasma and transfer to another test tube. Add 200 μL of methanol containing a predetermined internal standard to the plasma to precipitate plasma proteins. LC-MS / MS method is used to determine the plasma concentration of the compound.

[0118] In our previous studies, although δ-T3H was screened as a potential candidate for anti-radiation drugs, and it showed certain effects in the G-CSF induction experiment, it was found in the experiment that, unexpectedly, δ-T3H intravenous administration almost failed to show anti-radiation effects. At the same time, it was generally believed in past studies that δ-T3H has low bioavailability and a very short half-life, and is therefore not suitable for making anti-radiation drugs. At the same time, past studies have never involved research on improving the bioavailability of subcutaneous administration through structural modification, but generally achieved bioavailability improvement by adjusting the formulation. However, in the case where the bioavailability of δ-T3H is extremely low and intravenous administration cannot show anti-radiation effects, the improvement through the formulation method is likely to be extremely limited.

[0119] In this study, we attempted to modify δ-T3H and combine it with a specific route of administration to study the rules of improving absorption, distribution, metabolism and excretion (ADME) of δ-T3H ether derivatives, determine the pharmacokinetic characteristics of δ-T3H and selected derivatives in ICR mice, and improve their bioavailability. In our study, the pharmacokinetic characteristics of δ-T3H were first tested at a dose of 100 mg / kg by intravenous and subcutaneous injection, and the plasma concentration of mice was monitored for 24 hours after administration. The pharmacokinetic characteristics related to δ-T3H were calculated and listed in Table 1. Since the detection time of plasma concentration only lasted for 24 hours, it was not enough to calculate the t1 / 2 and AUC of subcutaneous injection. 0-∞ , CL and V dss It can be seen that δ-T3H not only fails to show anti-radiation effect after intravenous administration, but also has extremely low bioavailability after subcutaneous injection, and cannot meet the conditions for drug development at all.

[0120] Table 1 PK characteristics of δ-T3H and 8a in ICR mice (n = 3 / group)

[0121]

[0122]

[0123] In our study, δ-T3H and 8a were first tested at a dose of 100 mg / kg by intravenous injection and subcutaneous injection. The relevant plasma concentration-time curves were calculated and listed in Table 1. By subcutaneous injection, both δ-T3H and 8a showed slow distribution (the time to reach the maximum plasma concentration was 17.3 hours for δ-T3H and 12 hours for 8a). Compound 8a showed a higher plasma concentration than δ-T3H after subcutaneous injection (the maximum plasma concentration of 8a was 1142.23 ng / mL and that of δ-T3H was 495.01 ng / mL). Therefore, the subcutaneous injection AUC0-t value of 8a was significantly higher (7876.31 h·ng / mL for δ-T3H and 2.6×10 5 h·ng / mL for 8a). Since the plasma concentration was monitored for only 24 hours, the t1 / 2, AUC0-∞, CL, and Vdss after subcutaneous injection could not be calculated. The clearance rate of 8a was higher than that of δ-T3H. However, compared with δ-T3H, the subcutaneous bioavailability of 8a (F = 8.3%) seemed to be improved after chemical modification. By subcutaneous injection, a bioavailability increase of more than 400% was achieved. Overall, the results showed that δ-T3H ether derivatives (such as 8a) could greatly promote absorption and distribution without affecting metabolism. This design strategy unexpectedly greatly improved the bioavailability of active δ-T3H.

[0124] Peripheral blood recovery of δ-T3H and 8a after sub-lethal dose irradiation

[0125] Hematopoietic stem cells are sensitive to ionizing radiation. Damage to bone marrow hematopoietic stem cells caused by radiation leads to the loss of hematopoietic cells, which in turn triggers neutropenia, thrombocytopenia, and anemia. Therefore, δ-T3H and its derivative 8a were selected to further compare their radioprotective effects against radiation-induced cytopenia. Mice were randomly and equally divided into groups. Twenty-four hours before 6.5 Gy total body irradiation (TBI) (n = 8), they were respectively subcutaneously injected with PEG400 (solvent), δ-T3H (100 mg / kg), and 8a (100 mg / kg). At 0, 1, 4, 7, 10, 14, 18, 22, and 30 days after TBI, 20 μL of peripheral blood samples were collected from the tail vein and added to 2 mL of blood cell diluent. The samples were analyzed using a Celltac E fully automatic blood analyzer (Etda, Japan). Curves of peripheral cell counts of white blood cells (WBC), red blood cells (RBC), neutrophils, and platelets, as well as the curve of hemoglobin levels, were plotted.

[0126] As Figure 4 shown, the numbers of white blood cells, neutrophils, platelets, and red blood cells in the irradiated solvent group of mice decreased significantly and gradually; in contrast, δ-T3H and compound 8a significantly delayed the radiation-induced reduction. Both δ-T3H and 8a advanced the nadir days of neutrophils, platelets, and red blood cells, and simultaneously increased the nadir counts of neutrophils on day 7, platelets on day 10, and red blood cells on day 14. Combining the results of their survival advantages, it was shown that the pre-administration of 8a significantly improved the hematological recovery after radiation injury.

[0127] Recovery of hematopoietic stem cells and progenitors in the bone marrow with δ-T3H and 8a after sublethal dose irradiation

[0128] To investigate the in vivo effects of δ-T3H and its derivatives on promoting the recovery of HSPCs after irradiation and to compare their potencies, mice were subcutaneously injected with 100 mg / kg of δ-T3H and 8a before exposure to 6.5 Gy TBI. Ten days after TBI, all mice were sacrificed by cervical dislocation and immersed in 75% alcohol. The bilateral femurs were dissected, and the adherent muscle tissues were removed. Then, the femurs were rinsed with 1 mL of RPMI-1640 medium (Macgene, Beijing) containing 2% fetal bovine serum to harvest a single cell suspension of bone marrow (BM) cells.

[0129] As Figure 5 shown, consistent with the peripheral blood recovery data of mice irradiated with 6.5 Gy, the administration of 8a significantly increased the total bone marrow cell number compared to solvent-treated mice. Among BMNCs, hematopoietic progenitors (HPCs) and hematopoietic stem cells (HSCs) were classified as LK (Lin - Sca-1 - C-kit+ ) and LSK (Lin - Sca-1 + C-kit + ), the percentages and total numbers of LKs in 8a-treated and δ-T3H-treated mice were significantly increased on day 10 after TBI.

[0130] Next, according to the surface expression of CD34 and Flt3 (CD135), the cells in the LSK group were further divided into three subsets, namely the long-term hematopoietic stem cell (LT-HSCs) subset, the short-term hematopoietic stem cell (ST-HSCs) subset, and the multipotent progenitor cell (MPPs) subset. Compared with the solvent group, the frequency of LT-HSCs in the LSK group was significantly decreased, while the frequencies of LT-HSCs and MPPs in the LSK group were significantly increased in all δ-T3H- and 8a-treated mice.

[0131] Extramedullary hematopoiesis of δ-T3H and 8a in the spleen after sublethal irradiation

[0132] To compare and evaluate the induction effect of T3H and its derivative 8a on splenic EMH under radiation stress, mice were given a whole-body irradiation (TBI) of 6.5 Gy 24 hours after receiving a 100 mg / kg dose of these compounds. On day 10 after TBI, the expansion of splenic hematopoietic progenitor cells and stem cells (HPSCs) was evaluated by flow cytometry. As Figure 6 shown, the treatment with δ-T3H and 8a led to significant splenomegaly in irradiated mice and significantly increased the spleen coefficient. These results indicate that 8a can induce splenic EMH after radiation and promote the accelerated recovery of peripheral blood cells.

[0133] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A tocotrienol ether derivative, its isomer, its prodrug, its pharmaceutically acceptable salt, its hydrate or its solvate, characterized in that, The formula I is as follows, wherein R is selected from hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C6-C20 aryl, or substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl or heterocycloalkyl.

2. The tocotrienol ether derivative according to claim 1, wherein The said substitution means that one or more (preferably 1, 2, 3 or 4) hydrogen atoms on the group are each independently substituted by a substituent selected from the following group: halogen, -CN, -OR1, nitro, NR1R2, C1-C6 alkyl, halogenated C1-C4 alkyl (such as -CF3), C3-C8 cycloalkyl or heterocycloalkyl, C1-C6 alkylthio, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, C6-C12 aryl, 5-12 membered heteroaryl, COR3; wherein, R1 and R2 are each independently selected from H, C1-C4 alkyl; R3 is selected from NR1R2, OH, OR1.

3. The tocotrienol ether derivative according to claim 1, wherein R is selected from substituted C1-C6 alkyl; wherein, the said substitution means that one hydrogen atom on the group is substituted by OH.

4. The tocotrienol ether derivative according to claim 1, characterized in that, The said halogen substitution is by fluorine, chlorine, bromine or iodine.

5. The tocotrienol ether derivative according to claim 1, wherein R is selected from:

6. A composition comprising any one of the tocotrienol ether derivatives, isomers, prodrugs, pharmaceutically acceptable salts, hydrates or solvate compounds thereof as claimed in claims 1-5, and a pharmaceutical carrier or excipient.

7. The composition according to claim 6, wherein the composition is selected from gastrointestinal dosage forms and parenteral dosage forms.

8. A method for preparing the tocotrienol ether derivative according to claim 5, characterized in that, Comprising: The δ-T3H ester compounds 6a-e are obtained by a δ-T3H nucleophilic substitution reaction; or, The carboxylic acid derivatives 7a-e are obtained by hydrolyzing the ester compounds 6a-e; Or, The ester groups of the compounds 6a-e are reduced to hydroxyl groups to obtain the alcohol derivatives 8a-e; or, In the presence of KOH, δ-T3H methanesulfonate reacts with substituted ethanolamine to generate the target amine compounds 10a-c.

9. Use of the tocotrienol ether derivative according to any one of claims 1 - 5, its isomer, its prodrug, its pharmaceutically acceptable salt, its hydrate or its solvate, characterized in that, For (1) preparing anti-radiation drugs; (2) preparing drugs for inducing G-CSF production; (3) preparing drugs for peripheral blood recovery; (4) preparing drugs for restoring hematopoietic stem cells and progenitor cells in bone marrow; and / or (5) preparing drugs for promoting extramedullary hematopoiesis.

10. An active ingredient combination, the active ingredient combination comprising the following components: (1) The tocotrienol ether derivative, isomer, prodrug, pharmaceutically acceptable salt, hydrate or solvate compound thereof as claimed in claim 1; and (2) Anti-radiation drugs.

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