CN117915906A

CN117915906A - Methods for assessing patient response to treatment of neurodegenerative diseases with deuterated arachidonic acid

Info

Publication number: CN117915906A
Application number: CN202280021035.6A
Authority: CN
Inventors: P·米尔纳; M·S·什切皮诺夫
Original assignee: M SShiqiepinuofu; P Mierna; Retrotope Inc
Current assignee: M SShiqiepinuofu; P Mierna; Retrotope Inc
Priority date: 2021-02-05
Filing date: 2022-02-04
Publication date: 2024-04-19

Abstract

Diagnostic methods for assessing whether therapeutic concentrations of deuterated polyunsaturated fatty acids are reached in diseased neurons of a patient during treatment of neurodegenerative diseases are disclosed.

Description

Methods for assessing patient response to treatment of neurodegenerative diseases with deuterated arachidonic acid

Cross Reference to Related Applications

The present application claims priority from U.S. patent application Ser. No.17/169,271, U.S. patent application Ser. No.17/391,909, U.S. provisional application Ser. No.63/177,794, U.S. patent application Ser. No. 21, 4/2021, each of which is incorporated herein by reference in its entirety.

Technical Field

State of the art

Lipid autoxidation of polyunsaturated fatty acids (PUFAs) in neurons is associated with the pathology of many neurodegenerative diseases. The core of this oxidative pathway is the labile bis-allylic hydrogen atom found in arachidonic acid, the major PUFA in neurons.

In the cell membrane, arachidonic acid stacks together and oxidation processes involving Reactive Oxygen Species (ROS) act as initiators for the autoxidation of PUFAs by extracting the bis-allylic hydrogen from these PUFAs and forming oxidation-reactive species. Initial oxidation of the first bis-allylic site in the cell membrane or mitochondria results in more continuous oxidation of PUFAs. The oxidation process begins with hydrogen extraction at the bis-allylic site on the first PUFA, then proceeds continuously to the next PUFA, and so on. At some point, the oxidation process can damage or destroy the viability of neurons, resulting in further progression of disease conditions that produce excessive ROS.

To date, it has been disclosed in the art that progression of neurodegenerative disease conditions can be slowed by deuteration at one or more of the bis-allylic sites of arachidonic acid found in neurons. During this oxidation, the strength and stability of the carbon deuterium bond is significantly stronger than that of the carbon hydrogen bond. This means that carbon deuterium bonds can reduce the production of bis-allylic site oxidized species, thereby inhibiting the autoxidation pathway. Termination of this pathway, in turn, leads to increased survival of neurons, thus slowing progression of the disease.

Clinical studies to date have shown that certain neurodegenerative diseases respond to this treatment, including bovine head disease, amyotrophic Lateral Sclerosis (ALS), infant axonal dystrophy (INAD), and Friedreich's Ataxia (FA). Furthermore, preclinical studies have demonstrated that certain other neurodegenerative diseases respond similarly to this treatment, including the following examples: mitochondrial defects involving ROS, alzheimer's disease, huntington's disease, parkinson's disease, schizophrenia and bipolar disorders.

The manner in which these neurodegenerative diseases are treated with deuterated PUFAs is generally by administering active deuterated PUFAs or prodrugs thereof. For example, in neurodegenerative diseases, the active deuterated PUFA is deuterated arachidonic acid. However, such diseases may be treated by administration of deuterated arachidonic acid or a prodrug thereof. In one instance, the prodrug is a C ₁-C₅ alkyl ester of deuterated arachidonic acid. In another instance, the prodrug is deuterated linoleic acid or a C ₁-C₅ alkyl ester thereof. In the case of administration of any of these esters of PUFAs, they are readily removed by conditions in the stomach to provide the free acid or salt thereof. In the case of administration of deuterated linoleic acid, a portion of the compound is enzymatically converted to deuterated arachidonic acid as a source for providing deuterated arachidonic acid in the patient.

While such therapies have proven successful in clinical studies and animal models, it is necessary to determine whether therapeutic concentrations of deuterated arachidonic acid are reached or maintained in neurons of a particular patient. This concern arises from a variety of complications that are difficult to control. The method comprises the following steps:

PUFAs incorporated into cell membranes, including arachidonic acid, the half-life of which is determined by its turnover rate—the rate at which damaged PUFAs are cleared and replaced with new PUFAs, combined with the rate at which arachidonic acid molecules are extracted from cell membranes as part of bioconversion, including bioconversion to prostaglandins, prostacyclin (PGI ₂), thromboxane a ₂(TxA₂) or a series of hydroxyeicosatetraenoic acids (HETEs), epoxyeicosatrienoic acids (EETs), dihydroxyeicosatrienoic acids (DiHETEs), and other various catabolites;

the amount of polyunsaturated fatty acids (including deuterated PUFAs) taken up by humans depends on the total amount of polyunsaturated fatty acids taken up by humans. In general, human metabolism fails to absorb all of the PUFA consumed, and as the amount of PUFA consumed increases, the percentage of PUFA that is absorbed decreases. Since deuterated PUFAs are included in the total amount of PUFAs ingested, the percentage of non-absorbed PUFAs is equivalent to the percentage of non-absorbed deuterated PUFAs. Thus, if only 80% of the PUFA is absorbed, only 80% of the deuterated PUFA is absorbed. Further, the amount absorbed is a variable having individual differences;

The amount of PUFA absorbed also depends on whether this absorption is disturbed by digestion problems, such as diarrhea associated with clostridium difficile (c. In this case, much of the nutrition, including ingested PUFAs, is not absorbed into the body, but is merely "washed away";

even in specific individuals, their diet will not change at all. Thus, the uptake and percentage of absorption of PUFAs will change over time; and

Finally, the patient compliance with deuterated PUFA dosages is a factor related to the amount of deuterated PUFA absorbed.

All of these problems determine that patients receiving deuterated PUFA treatment should be evaluated to determine whether therapeutic concentrations are reached and/or maintained in their neurons.

The assessment of the concentration of deuterated arachidonic acid in neurons is more complex because neurons are inaccessible and the clinician cannot assess the concentration of deuterated arachidonic acid contained therein by taking a sample of these neurons. While the administered deuterated arachidonic acid can be systematically incorporated into all cells throughout the body, the data so far indicate that the body shunts most of the deuterated arachidonic acid into neurons and spinal fluid compared to other parts of the body. Thus, the concentration of deuterated arachidonic acid in replacement cells or reporter cells (e.g., skin cells, erythrocytes, etc.) is not necessarily representative of its concentration in neurons or cerebrospinal fluid. Furthermore, spinal puncturing to extract cerebrospinal fluid is not feasible because spinal puncturing requires a significant amount of time (e.g., about 30 minutes) and can cause significant discomfort to the patient, including headaches and back pain that last days or weeks.

Thus, there is a continuing need for a safe and effective diagnostic test to determine whether a neurodegenerative disease patient treated with a deuterated PUFA has a therapeutic or sub-therapeutic concentration of such deuterated arachidonic acid in its neurons. In addition, it helps to assess whether the patient's metabolism demonstrates proper absorption of deuterated arachidonic acid.

Disclosure of Invention

The present application provides a diagnostic test for diagnosing the presence or absence of therapeutic concentrations of deuterated arachidonic acid in neurons of a patient suffering from neurodegenerative diseases treated with deuterated arachidonic acid or a prodrug thereof without contacting the neurons or cerebrospinal fluid of the patient. The test is based in part on defining repeatable correlations for different clinical studies, all of which use the same deuterated arachidonic acid, ester thereof, or prodrug thereof to treat different classes of treatable neurodegenerative diseases.

The amount of deuterated arachidonic acid incorporated into the cells increases gradually over time. Thus, therapeutic concentrations of deuterated arachidonic acid cannot be immediately achieved. In contrast, there is a lag between the beginning of the treatment and the time the therapeutic outcome occurs, which can vary from patient to patient. At some point after the onset of treatment of neurodegenerative disease, one or more assessments of the patient's physical response to the treatment are required to determine when the treatment outcome is achieved and whether to maintain the treatment outcome. In one embodiment, this physical assessment is combined with obtaining a sample of patient-reported cells for determining the concentration of deuterated arachidonic acid in the cells. Or samples of patient reporter cells are obtained at regular intervals. When the patient's treatment outcome is first confirmed, the clinician or stationer can correlate the concentration of deuterated arachidonic acid in the then reported cells with the minimum concentration required for treatment.

When evaluating the therapeutic concentrations of clinical studies of different neurodegenerative diseases, the data support the conclusion that the therapeutic concentrations of deuterated arachidonic acid that produce therapeutic results for these different diseases are equal. Since each of these diseases involves a different etiology, it is surprising to find the fact that the concentration of deuterated arachidonic acid with therapeutic effect is equal.

Thus, in one embodiment, a method is provided for determining the minimum therapeutic concentration of deuterated arachidonic acid in neurons of patients suffering from different neurodegenerative diseases that can be treated with deuterated arachidonic acid without entering neurons or cerebrospinal fluid of said patients, comprising:

a) Obtaining data from a human clinical study for a representative number of neurodegenerative diseases, each of which can be treated with the same deuterated arachidonic acid or prodrug thereof, the data comprising the concentration of deuterated arachidonic acid in a reporter cell population measurable one or more times after initiation of treatment;

b) Assessing when the patient has evidence of treatment outcome;

c) Determining the average time from initiation of treatment to occurrence of the treatment outcome, and reporting the average concentration of deuterated arachidonic acid in the cells at the occurrence of the treatment outcome for each neurodegenerative disease;

d) Correlating the average concentration of deuterated arachidonic acid in the reporter cell population with the average time to onset of the minimum therapeutic outcome required for each neurodegenerative disease to demonstrate that the neurons have the minimum therapeutic concentration of deuterated arachidonic acid required for that disease;

e) Confirming that the minimum therapeutic concentration of said deuterated arachidonic acid and the average time for occurrence of the therapeutic outcome of each neurodegenerative disease are equal in other neurodegenerative diseases; and

F) The average minimum required therapeutic concentration and the average time at which the therapeutic result occurs are applied to other neurodegenerative diseases based on the equality.

In some embodiments, a non-invasive method is provided for determining the concentration of deuterated arachidonic acid in reporter cells of a patient population as a function of the time of occurrence of a therapeutic effect in a patient population having a neurodegenerative disease treatable with the deuterated arachidonic acid, the method comprising:

a) Obtaining data from a population of patients having a neurodegenerative disease for the first time being treated with deuterated arachidonic acid or a prodrug thereof, the data comprising the concentration of deuterated arachidonic acid obtained in a patient reporter cell population one or more times after initiation of treatment;

b) Assessing when a patient exhibits a therapeutic outcome;

c) Determining the average time from initiation of treatment to occurrence of the therapeutic outcome for each neurodegenerative disease and reporting the average concentration of deuterated arachidonic acid in the cells at the occurrence of the therapeutic outcome;

d) Correlating the average concentration of deuterated arachidonic acid in the patient report cell population with the average time to onset of the lowest desired therapeutic outcome for the neurodegenerative disease to demonstrate that the patient neurons have the lowest therapeutic concentration of deuterated arachidonic acid required for the disease;

e) Repeating steps a) -d) for one or more neurodegenerative diseases;

f) Assessing the minimum therapeutic concentration of deuterated arachidonic acid and the average time for occurrence of therapeutic outcomes for each of the one or more neurodegenerative diseases; and

F) An average minimum required therapeutic concentration for a group of neurodegenerative diseases and an average time for the occurrence of therapeutic results are obtained.

In some embodiments, a method is provided for determining a patient's response to administration of a composition comprising deuterated arachidonic acid or a prodrug thereof during treatment of a treatable neurodegenerative disease, the method comprising:

Predetermining when the outcome of the treatment of the disease should be apparent, obtaining a population of erythrocytes from a patient suffering from a neurodegenerative disease at the set point in time after initiation of the treatment;

Assessing the concentration of said deuterated arachidonic acid in said red blood cells at a time wherein said concentration is related to the content of arachidonic acid including deuterated arachidonic acid;

Comparing the estimated concentration to the predetermined concentration to determine when a treatment outcome is apparent;

Determining whether the estimated concentration is therapeutic or sub-therapeutic for a treatable neurodegenerative disease;

optionally, varying the dosage of deuterated arachidonic acid or a prodrug thereof administered to the patient according to the assay results.

In one embodiment, a method is provided for determining whether a patient receiving treatment of a therapeutic neurodegenerative disease with deuterated arachidonic acid or a prodrug thereof has reached the start of treatment or is maintaining a therapeutic concentration of said deuterated arachidonic acid in neurons of the patient, the method comprising:

Determining in advance when the outcome of the treatment of the disease should be apparent, obtaining a population of reporter cells from a patient suffering from a neurodegenerative disease at the set point in time after initiation of the treatment;

Assessing the concentration of deuterated arachidonic acid in the reporter cell at the time;

Determining whether the estimated concentration is therapeutic or sub-therapeutic for a treatable neurodegenerative disease; and

In one embodiment, the method is repeated thereafter to assess whether the patient is reaching or maintaining the therapeutic concentration.

In one embodiment, the sample of reporter cells is obtained at a time period set to about 1 month after initiation of treatment. Optional subsequent samples may be obtained at intervals, for example, 1 month thereafter, 3 months thereafter, half a year or once a year.

In one embodiment, the deuterated arachidonic acid is D2-arachidonic acid, D4-arachidonic acid, or D6-arachidonic acid. Examples of D2-arachidonic acid include, but are not limited to, 7-D2-arachidonic acid, 10-D2-arachidonic acid, and 13, 13-D2-arachidonic acid. Examples of D4-arachidonic acid include, but are not limited to 7,7,10,10-arachidonic acid, 7,7,13,13-D4-arachidonic acid, 10,10,13,13-D4-arachidonic acid. D6-arachidonic acid (as defined herein) includes 7,7,10,10,13,13-D6-arachidonic acid. Preferably, the deuterated arachidonic acid is D2-arachidonic acid or D6-arachidonic acid. In some embodiments, D6-arachidonic acid comprises both 7,7,10,10,13,13-D6-arachidonic acid and a composition of deuterated arachidonic acid, at least 90% of the hydrogen atoms at the 7,10, 13 positions of the composition being replaced by deuterium, and having at least one non-bis-allyl position with a level of deuteration that is greater than its natural abundance. In one embodiment, the non-bis-allylic positions optionally have up to about 35% of the remaining hydrogen atoms substituted with deuterium, and preferably, about 1 to 35%, more preferably, about 1 to 10%.

In one embodiment, the reporter cell is any accessible cell in the body and preferably is easily accessible without undue pain or inconvenience to the patient. For example, suitable reporter cells include only erythrocytes, skin cells, adipocytes, biopsy cells, including epithelial cells found in urine, and the like. In one embodiment, the reporter cell is preferably a red blood cell.

In one embodiment, a method is provided for determining a patient's response to administration of a composition comprising deuterated arachidonic acid or a prodrug thereof during treatment of a treatable neurodegenerative disease, the method comprising:

Assessing the concentration of said deuterated arachidonic acid in said erythrocytes at that time;

comparing the estimated concentration to the predetermined concentration to determine when a therapeutic result is apparent;

In one embodiment, clinical studies of each of the treatable neurodegenerative diseases demonstrate that the concentration of deuterated 13, 13-D2-arachidonic acid (about 3% or more relative to the total amount of arachidonic acid including deuterated arachidonic acid) in erythrocytes as reporter cells correlates with the minimum concentration required to achieve therapeutic results in these neurodegenerative diseases. Thus, this concentration establishes a baseline for the treatment of neurodegenerative diseases that can be treated with 13, 13-D2-arachidonic acid, and evidence suggests that the concentration of 13, 13-D2-arachidonic acid in diseased neurons is sufficient to provide treatment for treatable diseases.

In one embodiment, the invention provides a diagnostic test for determining whether the concentration of deuterated 13, 13-D2-arachidonic acid is therapeutic or sub-therapeutic in a patient having a treatable neurodegenerative disease without contacting neurons or cerebrospinal fluid of the patient, comprising:

obtaining a blood sample from a patient receiving 13, 13-D2-arachidonic acid or an ester thereof for treatment of a neurodegenerative disease;

Determining the concentration of 13, 13-D2-arachidonic acid in Red Blood Cells (RBCs); and

This concentration is compared to the minimum therapeutic concentration of 13, 13-D2-arachidonic acid based on at least 3% of the total amount of arachidonic acid including deuterated arachidonic acid in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D2-arachidonic acid in the neurons.

In one embodiment, a higher concentration of 13, 13-D2-arachidonic acid is believed to enhance the therapeutic outcome, and the attending clinician may conclude that such higher concentration should be the target concentration for a particular patient or group of patients to produce the therapeutic outcome. For example, the concentration of 13, 13-D2-arachidonic acid may be set to 4%,5%,6%,7%,8%,9% or even 10% as a specific therapeutic goal for a given patient, recognizing that the concentration of 13, 13-D2-arachidonic acid is the baseline for the lowest desired therapeutic outcome. The attending clinician may increase the target concentration based on the age and weight of the patient, the condition of the patient, the progress of the disease, and other factors known to the clinician.

The minimum therapeutic concentrations of D2-arachidonic acid provided above may be used to correlate the minimum therapeutic concentrations of D4-arachidonic acid or D6-arachidonic acid, recognizing that deuteration at one or both of the remaining bis-allylic sites increases the stability of arachidonic acid to lipid autoxidation.

In one embodiment, a diagnostic test is provided to determine whether the concentration of deuterated D4-arachidonic acid is therapeutic or sub-therapeutic in a patient with a treatable neurodegenerative disease without contacting neurons or cerebrospinal fluid of the patient, the test comprising:

obtaining a blood sample from a patient suffering from a neurodegenerative disease being treated with D4-arachidonic acid or an ester thereof;

Determining the concentration of D4-arachidonic acid in Red Blood Cells (RBCs); and

This concentration is compared to the minimum therapeutic concentration of D4-arachidonic acid based on at least 1% of the total amount of arachidonic acid including deuterated arachidonic acid in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D4-arachidonic acid in the neurons.

In one embodiment, a diagnostic test is provided to determine whether the concentration of deuterated D6-arachidonic acid in a patient with a treatable neurodegenerative disease is therapeutic or sub-therapeutic without contacting neurons or cerebrospinal fluid of the patient, the test comprising:

Obtaining a blood sample from a patient with a neurodegenerative disease treated with D4-arachidonic acid or an ester thereof;

Determining the concentration of D6-arachidonic acid in Red Blood Cells (RBCs); and

This concentration is compared to a minimum therapeutic concentration of D6-arachidonic acid of at least 0.5% based on the total amount of arachidonic acid including deuterated arachidonic acid in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D6-arachidonic acid in the neurons.

In one embodiment, higher concentrations of D4-or D6-arachidonic acid may enhance the therapeutic outcome, and the clinician may conclude that if the enhanced outcome is set to the target concentrations, these concentrations are the lowest target concentrations required for the therapeutic outcome to occur. For example, the concentrations of D4-and D6-arachidonic acid can be set as in Table 1, wherein the concentrations are determined as follows:

TABLE 1

* In erythrocytes

In one embodiment, a kit is provided that includes diagnostic materials for performing the method and a correlation table correlating the concentration of deuterated arachidonic acid in red blood cells to the concentration of arachidonic acid in cerebrospinal fluid of a human. In addition to the correlation table, such diagnostic materials may include one or more instructions as to when such testing should be performed, factors suggesting delay of such testing, and the like.

Drawings

FIG. 1 shows a graph of the percentage of 13, 13-D2-arachidonic acid in Red Blood Cells (RBC) and Spinal Fluid (SF) at a given time point after onset of treatment with 11, 11-D2-linoleic acid in adult patients.

FIG. 2 shows a graph of the percentage of 13, 13-D2-arachidonic acid in Red Blood Cells (RBC) and Spinal Fluid (SF) at a given time point after onset of treatment with 11, 11-D2-linoleic acid in an underage patient.

Detailed Description

A diagnostic method or test is provided for determining whether a minimum therapeutic concentration of deuterated arachidonic acid is present in neurons of a patient undergoing treatment for neurodegenerative disease. Methods or tests are also provided to determine whether a patient receiving a treated neurodegenerative disease has reached or is maintaining a therapeutic concentration of deuterated arachidonic acid.

Before discussing the present invention in more detail, the following terms are first defined. Undefined terms give their definitions in the context or give a medically acceptable definition.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

As used herein, the term "about" when used before the numerical designations of other inclusive ranges, such as temperature, time, amount, concentration, etc., means that the approximation may be plus (+) or minus (-) 10%, 5%, 1%, or any subrange or sub-value therebetween. Preferably, the term "about" when used in reference to the lowest therapeutic concentration means a dosage variation of +/-10%.

As used herein, the terms "comprising" or "comprises" are intended to mean that the compositions and methods include the listed elements, but not exclude other elements.

As used herein, the term "consisting essentially of … …" when used in defining compositions and methods shall mean excluding other elements having any substantial significance to the combination of objects. Thus, a composition consisting essentially of the elements defined herein does not exclude other materials or steps that do not materially affect the basic and novel characteristics of the claimed invention.

As used herein, the term "consisting of … …" shall mean other ingredients and substantial process steps that exclude more than trace elements. Embodiments defined by each of these transitional terms are within the scope of the present invention.

As used herein, the term "linoleic acid" refers to compounds having the formula:

Linoleates are formed by substitution of the-OH group with-OR. These esters are defined below.

As used herein, unless the context indicates otherwise, the term "deuterated linoleic acid or an ester thereof refers to a compound of linoleic acid or an ester thereof that contains one or two deuterium atoms at its 11-position, and optionally additional deuterium atoms at other positions within the molecule (including the 8-position). Specific compounds encompassed by this definition include, by way of example only, 11-D1-linoleic acid, 11-D2-linoleic acid, 8, 11-D2-linoleic acid, 8,11,11-D3-linoleic acid, and 8,8,11,11-D4-linoleic acid, as well as esters of any of these compounds. Additional stabilization of the bis-allylic site may also include replacement of one or more bis-allylic carbon atoms with a heavy isotope alone or in combination with deuteration (or tritiation), as Isotope Effects (IEs) result in stabilization of the bond with the heavy isotope being additive according to basic chemistry established over time. (WESTHEIMER, chem. Rev. (1961), 61:265-273;Shchepinov,Rejuvenation Res, (2007), 10:47-59; hill et al, free radio. Biol. Med., (2012), 53:893-906; andrey ev et al, free radio. Biol. Med., (2015), 82:63-72.Bigeleisen, j. Use tracers to track the effectiveness of chemical reactions (THE VALIDITY of the use of tracers to follow chemical reactions.) science, (1949), 110:14-16.

As used herein, arachidonic acid has a numbering system as described below:

wherein each of positions 7, 10 and 13 is a bis-allyl site within the structure.

As used herein, unless the context indicates otherwise, the term "deuterated arachidonic acid or ester thereof" refers to an arachidonic acid or ester compound having at least one deuterium atom at the bis-allyl position, and optionally additional deuterium atoms at other positions within the molecule. The deuterated arachidonic acid includes monoallyl deuterated arachidonic acid having a deuterium atom at the 7, 10, or 13 position and di, tri, tetra, penta, hexadeuteration at the diallyl position and optionally deuteration at a level greater than the natural abundance of deuterium at positions other than the diallyl position.

The term deuterated arachidonic acid includes 7, 7-D2-arachidonic acid, 10-D2-arachidonic acid, and 13, 13-D2-arachidonic acid. The term D4-arachidonic acid includes 7,7,10,10-D4-arachidonic acid, 7,7,13,13-D4-arachidonic acid, 10,10,13,13-D4-arachidonic acid. The term D6-arachidonic acid includes 7,7,10,10,13,13-D6-arachidonic acid as well as deuterated arachidonic acid having at least 90% of the hydrogen atoms replaced by deuterium at the 7,10,10,13,13 positions and optionally up to 35% of the remaining hydrogen atoms at the non-diallyl positions replaced by deuterium.

As used herein, the term "ester" refers to any pharmaceutically acceptable ester of deuterated linoleic acid or deuterated arachidonic acid, such as, but not limited to, C1-C6 alkyl esters, glycerol (including mono-, di-and tri-glycerides), sucrose esters, phosphate esters, and the like. The particular ester employed is not critical as long as the ester is pharmaceutically acceptable (non-toxic and biocompatible).

As used herein, the term "prodrug of deuterated arachidonic acid" refers to esters of deuterated arachidonic acid (e.g., C1-C5 alkyl esters), deuterated linoleic acid, and esters of deuterated linoleic acid (e.g., C1-C5 alkyl esters). Esters of deuterated arachidonic acid and deuterated linoleic acid are readily converted to the corresponding acid/salt forms in the gastrointestinal tract after administration. A portion of the administered deuterated linoleic acid is enzymatically converted to deuterated arachidonic acid, thereby acting as a prodrug of such deuterated arachidonic acid. In one embodiment, 11-D2-linoleic acid is converted to 13, 13-D2-arachidonic acid. In one embodiment 8,8,11,11-D4-linoleic acid is converted to 10,10,13,13-D4-arachidonic acid.

The term "treatable neurodegenerative disease" or "neurodegenerative disease treatable with deuterated arachidonic acid" refers to a specific neurodegenerative disease that is considered treatable with deuterated arachidonic acid as compared to a neurodegenerative disease that cannot be treated with deuterated arachidonic acid. Neurodegenerative diseases that can be treated with deuterated arachidonic acid include Alzheimer's disease, mild cognitive impairment, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, friedel-crafts ataxia, parkinson's disease, tauopathies (including PSP) and Huntington's disease. On the other hand, the data to date indicate that the following diseases cannot be treated with deuterated arachidonic acid and/or other deuterated polyunsaturated fatty acids, including Tay-Sach's disease, GPX-4 deficiency and neuroserine.

As used herein, the term "etiology" refers to the cause of the disease. The term "pathogenesis" or "pathology" refers to the development, structural/functional changes, and natural course of the disease associated with the disease. The term "course of disease" refers to the progression of the disease without treatment or without treatment with deuterated arachidonic acid or a prodrug thereof.

As used herein, the term "rate of decrease in disease progression" refers to a decrease in the rate of disease progression after initiation of treatment as compared to the natural course of the patient. In one embodiment, the rate of decrease in disease progression in ALS is measured by determining the rate of disease progression during the course of the natural course using ALSFRS-R score, which is then measured again within a treatment interval after the start of treatment and at the end of a set period of time thereafter (e.g., 6 months). These two rates were then calculated annually, and the rate of disease progression reduction resulted in a percentage change of at least 30% before and after ALSFRS-R scoring.

For PSP, the rate of disease progression is measured by using a progressive supranuclear palsy rating scale (Progressive Supranuclear PALSY RATING SCALE) or a unified parkinson's disease rating scale (Unified Parkinson' S DISEASE RATING SCALE) to determine the rate of disease progression during the course of the natural disease, and again measure any score (e.g., 1 month and every 3 months thereafter) within the treatment interval after initiation of treatment and within a set time thereafter. These two rates were then calculated annually, with the rate of disease progression decreasing resulting in a percentage change of at least 30% from before and after scoring.

Likewise, there is a similar standardized scale to measure the rate of disease progression in Alzheimer's disease, parkinson's disease, huntington's disease, friedel-crafts ataxia and the like.

As used herein, the term "minimal desired therapeutic outcome" refers to a patient receiving treatment with deuterated arachidonic acid or a prodrug thereof that demonstrates a reduction in the rate of disease progression of at least about 30% as compared to the rate of disease progression determined during the course of the natural course.

As used herein, the term "patient" refers to a human patient or group of human patients suffering from a neurodegenerative disease. When more than 1 patient is evaluated, then the average of their disease progression is used.

As used herein, the term "pharmaceutically acceptable salts" of the compounds disclosed herein includes within the scope of the present invention acid or base salts that retain the desired pharmacological activity and are not biologically undesirable (e.g., the salts are not overly toxic, sensitized or irritating, and are bioavailable). When the compounds of the present invention have a basic group, such as an amino group, pharmaceutically acceptable salts may be formed from inorganic acids (e.g., hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid), organic acids (e.g., alginates, formic acid, acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, and p-toluenesulfonic acid), or acidic amino acids (e.g., aspartic acid and glutamic acid). When the compound of the present invention has an acidic group, such as a carboxylic acid group, it may form a salt with a metal (e.g., a base and an earth alkali metal (e.g., na+, li+, k+, ca2+, mg2+, zn2+), ammonia or an organic amine (e.g., dicyclohexylamine, trimethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine) or a basic amino acid (e.g., arginine, lysine, and ornithine).

Composition and method for producing the same

A portion of the deuterated linoleic acid or an ester thereof administered to a patient is converted in vivo as a deesterified acid or a salt thereof to deuterated arachidonic acid, and the deuterated linoleic acid portion so converted acts as a prodrug of deuterated arachidonic acid. There are a number of deuterated linoleic acid compounds that are commercially available or known in the art. These include, by way of example only, 11-D1-linoleic acid, 11-D2-linoleic acid, 8,8,11,11-D4-linoleic acid (which upon enzymatic desaturation extension yields 10,10,13,13-D4-arachidonic acid). Other deuterated linoleic acids are described in U.S. patent No. 10,052,299, incorporated herein by reference in its entirety. In addition, 11-D1-linoleic acid is available from Cayman chemical company (Anaba 48108, michigan, U.S.A.).

In addition to deuterated linoleic acid or an ester thereof, the enzymatic conversion required for the prodrug can be bypassed by administering deuterated arachidonic acid to the patient. Such deuterated arachidonic acid includes 7, 7-D2-arachidonic acid, 10-D2-arachidonic acid, 13-D2-arachidonic acid, 7,7,10,10-D4-arachidonic acid, 7,7,13,13-D4-arachidonic acid, 10,10,13,13-D4-arachidonic acid, 7,7,10,10,13,13-D6-arachidonic acid disclosed by Shchepinov et al Molecules,28 (12): 3331et seq. (2018), which is incorporated herein by reference in its entirety. Other deuterated arachidonic acid compounds are known in the art.

Still further, deuterated arachidonic acid may be prepared by a catalytic process as described in U.S. patent No. 10,577,304, which is incorporated herein by reference in its entirety. These procedures provide for essentially complete deuteration at the bis-allylic position, partial deuteration at the non-bis-allylic position and predominantly at the mono-allylic position. In general, deuteration at 3 bis-allylmethylene sites converts 6 hydrogen atoms to deuterium with greater than 90% efficiency—meaning that in the deuterated arachidonic acid group, 6 bis-allylhydrogen atoms have on average greater than 5.4 deuterium atoms. In addition, no more than about 35% (excluding any ester moieties) of the total number of hydrogen atoms in arachidonic acid are replaced with deuterium. Since there are 32 hydrogen atoms in arachidonic acid, the percentage of deuterium in these compounds ranges from about 15% to no more than 35%. It is understood that when the term "deuterated arachidonic acid" is defined to mean arachidonic acid D6, it includes arachidonic acid having on average more than 5.4 deuterium atoms at the bis-allyl position, and deuteration ranges from about 15% to no more than 35% based on the number of hydrogen atoms present on non-deuterated arachidonic acid, including protons on carboxylic acids.

The esters of these deuterated fatty acids are prepared by conventional techniques well known in the art. The esters are preferably derived from C1-C5 alkyl alcohols.

Method-deuterated linoleic acid

Some of the methods described herein utilize enzymatic conversion of deuterated linoleic acid or an ester thereof to provide deuterated arachidonic acid. In particular, it is well known that a fraction of linoleic acid or an ester thereof consumed by an individual is converted in vivo to arachidonic acid, whereby the fraction of linoleic acid or an ester thereof acts as a prodrug of arachidonic acid.

In one embodiment, a patient treated with 11, 11-D2-linoleic acid or an ester thereof will produce 13, 13-D2-arachidonic acid in vivo. As described above, the concentration of the 13, 13-D2-arachidonic acid in erythrocytes is considered therapeutic when it reaches at least about 3% based on the total number of arachidonic acid+deuterated arachidonic acid found therein.

In one embodiment, a patient treated with 8,8,11,11-D4-linoleic acid or an ester thereof will produce 10,10,13,13-D4-arachidonic acid in vivo. As described above, the concentration of 10,10,13,13-D4-arachidonic acid in erythrocytes is considered therapeutic when it reaches at least about 1% based on the total number of arachidonic acid+deuterated arachidonic acid found therein.

In one embodiment, a concentration of D6-arachidonic acid in erythrocytes is considered therapeutic when it reaches at least about 1% based on the total number of arachidonic acid+deuterated arachidonic acid found therein.

Surprisingly, each treatable neurodegenerative disease requires an equivalent deuterated arachidonic acid threshold to achieve the minimum therapeutic concentration required for therapeutic effect, even though each disease has its own etiology and, in many cases, its own pathology.

In one embodiment, deuterated linoleic acid, such as 11, 11-D2-linoleic acid, is administered in a dosing regimen comprising an initiating amount and a maintenance amount. In one embodiment, the initiator amount comprises periodic administration of 11, 11-D2-linoleic acid or an ester thereof. In one embodiment, the initiator amount comprises at least about 7 grams 11, 11-D2-linoleic acid or an ester thereof per day. In one embodiment, the amount of initiator is at least about 30 days, for example, due to enzymatic conversion of a portion of the 11, 11-D2-linoleic acid to 13, 13-D2-arachidonic acid, thereby rapidly achieving therapeutic concentrations of 13, 13-D2-arachidonic acid in vivo. In one embodiment, a maintenance dose is administered periodically after completion of the initiator dose, and the maintenance dose is at least about 3 grams 11, 11-D2-linoleic acid or ester thereof per day.

Deuterated arachidonic acid

Some of the methods described herein use deuterated arachidonic acid or an ester thereof. The method preferably employs D2-arachidonic acid, D4-arachidonic acid or D6-arachidonic acid as the specific deuterated arachidonic acid. In the case of D6-arachidonic acid, this includes the catalytically produced deuteration as described above. In any event, the concentration of deuterated arachidonic acid found in erythrocytes to have therapeutic effect is listed in table 1 above.

In one embodiment, deuterated arachidonic acid, such as 7,7,10,10,13,13-D6-arachidonic acid, is administered in a dosing regimen that includes an initiating and maintenance dose. In one embodiment, the initiator amount comprises periodic administration of 7,7,10,10,13,13-D6-arachidonic acid or an ester thereof. In one embodiment, the initiator amount comprises from about 0.5 grams to about 5 grams 7,7,10,10,13,13-D6-arachidonic acid or an ester thereof per day. In one embodiment, the amount of initiator is maintained for about 24 days to about 45 days, for example, to achieve a therapeutic concentration of 7,7,10,10,13,13-D6-arachidonic acid in vivo rapidly. In one embodiment, the maintenance dose is administered periodically after the priming dose is completed. In one embodiment, up to about 70% of the initiator amount of 7,7,10,10,13,13-D6-arachidonic acid or ester thereof is administered daily.

Detailed description of the preferred embodiments

Embodiment 1. A non-invasive method for determining the concentration of deuterated arachidonic acid in reporter cells of a patient population, the concentration being correlated with the occurrence of a therapeutic effect in a patient population having a neurodegenerative disease treatable with said deuterated arachidonic acid, the method comprising:

a) Obtaining data from a patient population first suffering from a neurodegenerative disease treated with deuterated arachidonic acid or a prodrug thereof comprising the concentration of deuterated arachidonic acid obtained in a patient reporter cell population one or more times after initiation of treatment;

b) Assessing when a patient exhibits a therapeutic outcome;

e) Repeating steps a) -d) for one or more neurodegenerative diseases;

Embodiment 2. A method for determining a patient's response to administration of a composition comprising deuterated arachidonic acid or a prodrug thereof during treatment of a treatable neurodegenerative disease, the method comprising:

assessing the concentration of said deuterated arachidonic acid in said erythrocytes at that time, wherein said concentration is relative to the amount of arachidonic acid present, including deuterated arachidonic acid;

Embodiment 3. The sample of reporter cells as in any one of the preceding embodiments, wherein the sample of reporter cells is obtained about 1 month after initiation of the treatment.

Embodiment 4. The method of any of the preceding embodiments, wherein further samples of reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

Embodiment 5. Any of the preceding embodiments, wherein the reporter cell is a red blood cell, a skin cell, an adipocyte, a biopsy cell, or an epithelial cell.

Embodiment 6. Any of the preceding embodiments, wherein the deuterated arachidonic acid is 13, 13-D2-arachidonic acid.

Embodiment 7. Any of the preceding embodiments, wherein the deuterated arachidonic acid is D4-arachidonic acid.

Embodiment 8. Any of the foregoing embodiments wherein the deuterated arachidonic acid is D6-arachidonic acid wherein the deuterated D6-arachidonic acid comprises at least 90% of hydrogen atoms replaced with deuterium at position 7,7,10,10,13,13 and optionally up to 35% of the remaining hydrogen atoms at non-bis-allyl positions are replaced with deuterium.

Embodiment 9. A method for assessing a patient's response to administration of a composition comprising deuterated arachidonic acid or a prodrug thereof during treatment of a treatable neurodegenerative disease, the method comprising:

assessing the concentration of deuterated arachidonic acid in red blood cells of the patient relative to the total amount of arachidonic acid present, including deuterated arachidonic acid;

Comparing the concentration to a predetermined minimum therapeutic concentration of deuterated arachidonic acid in the erythrocytes;

When the concentration of said deuterated arachidonic acid in the erythrocytes of the patient is lower than said determined therapeutic concentration, it is considered a sub-therapeutic concentration; when the concentration of deuterated arachidonic acid is equal to or greater than the determined therapeutic concentration, it is considered a therapeutic concentration; and

Optionally, the method is repeated as necessary to assess whether the patient's concentration of deuterated arachidonic acid in the erythrocytes has reached or is maintaining the therapeutic concentration.

Embodiment 10. Embodiment 9 wherein the reporter cell sample is obtained about 1 month after initiation of treatment.

Embodiment 11. Any of the preceding embodiments, wherein further samples of reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

Embodiment 12. Any of the preceding embodiments, wherein the reporter cell is a red blood cell, a skin cell, an adipocyte, a biopsy cell, or an epithelial cell.

Embodiment 13. Any of the preceding embodiments, wherein the deuterated arachidonic acid is 13, 13-D2-arachidonic acid

Embodiment 14. The method of any of the preceding embodiments, wherein the deuterated arachidonic acid is D4-arachidonic acid.

Embodiment 15. As in any of the preceding embodiments, wherein the deuterated arachidonic acid is D6-arachidonic acid wherein the deuterated D6-arachidonic acid comprises at least 90% of hydrogen atoms replaced with deuterium at position 7,7,10,10,13,13 and optionally up to 35% of the remaining hydrogen atoms at non-bis-allylic positions are replaced with deuterium.

Embodiment 16. A diagnostic method for determining whether the concentration of deuterated 13, 13-D2-arachidonic acid is therapeutic or sub-therapeutic in a patient suffering from a treatable neurodegenerative disease without intervention in neurons or cerebrospinal fluid of the patient, the method comprising:

Obtaining a blood sample from a patient receiving a neurodegenerative disease treated with 13, 13-D2-arachidonic acid or an ester thereof;

Comparing the concentration to a minimum therapeutic concentration of 13, 13-D2-arachidonic acid that is at least 3% of the total amount of arachidonic acid (including deuterated arachidonic acid) present in the red blood cells to determine whether the patient has a therapeutic or sub-therapeutic concentration of 13, 13-D2-arachidonic acid in the neurons.

Embodiment 17. The therapeutic concentration of 13, 13-D2-arachidonic acid as described in embodiment 16 is set to 4%, 5%, 6%, 7%, 8%, 9% or even 10% as therapeutic target for a given patient or group of patients.

Embodiment 18. Any of the preceding embodiments, wherein the reporter cell sample is obtained about 1 month after initiation of the treatment.

Embodiment 19. The method of any of the preceding embodiments, wherein further samples of reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

Embodiment 20. A diagnostic method for determining whether the concentration of deuterated D4-arachidonic acid in a patient suffering from a treatable neurodegenerative disease is therapeutic or sub-therapeutic without intervention in neurons or cerebrospinal fluid of the patient, the method comprising:

Obtaining a blood sample from a patient undergoing treatment for a neurodegenerative disease with D4-arachidonic acid or an ester thereof; and

This concentration is compared to the minimum therapeutic concentration of D4-arachidonic acid based on at least 1% of the total amount of arachidonic acid (including deuterated arachidonic acid) in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D4-arachidonic acid in the neurons.

Embodiment 21. The therapeutic concentration of D4-arachidonic acid is set to 2%, 3% or 5% as therapeutic target for a given patient or group of patients as in embodiment 20.

Embodiment 22. The sample of reporter cells is obtained about 1 month after initiation of treatment, as in any of the preceding embodiments.

Embodiment 23. Any of the preceding embodiments, wherein further samples of reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

Embodiment 24. A diagnostic method for determining whether the concentration of deuterated D6-arachidonic acid in a patient suffering from a treatable neurodegenerative disease is therapeutic or sub-therapeutic without intervention in neurons or cerebrospinal fluid of the patient, the method comprising:

This concentration is compared to the minimum therapeutic concentration of D6-arachidonic acid of at least 0.5% based on the total amount of arachidonic acid (including deuterated arachidonic acid) in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D6-arachidonic acid in the neurons.

Embodiment 25. Embodiment 24 wherein the therapeutic concentration of D6-arachidonic acid is set to 1%, 1.5%, 2%, 2.5% or 3% as a therapeutic target for a given patient or group of patients.

Embodiment 26. Any of the preceding embodiments, wherein the reporter cell sample is obtained about 1 month after initiation of the treatment.

Embodiment 27. Any of the preceding embodiments, wherein further samples of reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

Embodiment 28. As in any of the preceding embodiments, wherein the deuterated arachidonic acid is D6-arachidonic acid wherein the deuterated D6-arachidonic acid comprises at least 90% of hydrogen atoms replaced with deuterium at position 7,7,10,10,13,13 and optionally up to 35% of the remaining hydrogen atoms at non-bis-allylic positions are replaced with deuterium.

Embodiment 29. A kit comprising any of the preceding embodiments, wherein the diagnostic material for performing the method is included, and a correlation table correlating concentration of deuterated arachidonic acid in red blood cells to its concentration in human cerebrospinal fluid. In addition to the correlation table, such diagnostic materials may include one or more instructions regarding when such testing should be performed, factors suggesting delay of such testing, and the like.

Examples

The invention may be further understood by reference to the following examples, which are intended to be merely illustrative of the invention. The scope of the invention is not limited by the example embodiments, which are intended as illustrations of only a single aspect of the invention. Any functionally equivalent method is within the scope of the invention. Various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims. In these embodiments, the following terms are used herein and have the following meanings. If undefined, the abbreviation has its conventional medical meaning.

D2-aa=13, 13-D2-arachidonic acid

Aa=arachidonic acid

ALSFRS-R = revised ALS function rating scale

BID = twice a day

CNS = central nervous system

D2-la=11, 11-D2-linoleic acid

La=linoleic acid

PK = pharmacokinetics

PSP = progressive supranuclear palsy

RBC = red blood cell

SAE = severe adverse event

SF = cerebrospinal fluid

TID = three times a day

% Deuterated AA = percentage of deuterated arachidonic acid to total arachidonic acid (including deuterated arachidonic acid) in the cells analyzed

Example 1 determination of AA concentration in erythrocytes and spinal fluid/neurons in individual patients

The present example determines the relative concentrations of D2-AA in SF and RBC to determine if there is 1 between the two concentrations based on the relative amounts of D2-AA and D2-LA: 1. Specifically, 9 grams of ethyl D2-linoleate per day was continuously provided to the patient over a period of about six months. Blood and SF samples were collected periodically and the concentrations of D2-LA and D2-AA in the red blood cells and SF were measured. In all cases, D2-AA is obtained by deacylation of ethyl linoleate in the gastrointestinal tract, followed by conversion of D2-LA to D2-AA in vivo.

TABLE 2

The results in the table show that the concentration of D2-AA in cerebrospinal fluid has reached 8% calculated on the amount of arachidonic acid + deuterated arachidonic acid.

Next, table 3 shows the concentrations of D2-LA and D2-AA in erythrocytes at 3 months and 6 months in the same patient.

TABLE 3 Table 3

Note that the concentration of D2-AA in erythrocytes was significantly lower at 3 months than at 6 months, indicating an increase in D2-AA over time. Furthermore, the ratio of D2-LA to D2-AA was changed from 2.9:1 at 3 months to 2.1:1 at 6 months.

Since the amount of D2-AA increases in an incremental manner over time based on the conversion of D2-LA, which limits the amount of D2-AA produced biologically per day, a fairly linear rate of increase can be assumed. As shown in FIG. 1, where the solid line is set by the concentration of D2-AA at 3 months and 6 months, then extrapolated back to the beginning of treatment (0 months). The D2-AA value in erythrocytes at 1 month was estimated from this linear relationship. The values shown in SF at 1 month (circles) are also provided.

Based on the above and FIG. 1, it can be seen that the data to date show that the amount of D2-AA in erythrocytes at 1 month is about 3% and that the amount of D2-AA in SF is 8%. Thus, these data indicate that the human body shunts more AA (including D2-AA) into SF (and neurons) than erythrocytes and possibly other reporter cells.

Example 2-determination of AA concentration in erythrocytes and spinal fluid/neurons in a group of 14 patients

The concentration of D2-AA in erythrocytes was also determined in this example. Specifically, 14 children were provided with 3 grams of ethyl D2-linoleate per day for __ months, and then 2 grams of ethyl D2-linoleate per day for the remaining six months. Blood samples were taken from children other than 1 child at 3 months and from all children at 6 months. The concentration of D2-AA in the erythrocytes was measured. In all cases, D2-AA is obtained by deacylation of ethyl linoleate in the gastrointestinal tract, followed by conversion of D2-LA in vivo to D2-AA.

At 3 months, the average concentration of D2-AA in erythrocytes was determined to be 12% (as low as 6.8% and as high as 16.8%). At 6 months, the average concentration of D2-AA in erythrocytes was determined to be 16.7% (as low as 12.0%, as high as 26.1%). FIG. 2 depicts the results assuming that D2-AA accumulates in vivo in a linear relationship. Included in fig. 2 is data for D2-AA at 1 month in spinal fluid found in example 1.

It can be seen that figures 1 and 2 are substantially identical, which strongly suggests that D2-LA administration to adult patients in example 1 and children in example 2 maximizes D2-LA conversion to D2-AA. The data further indicate that once maximized, the amount of D2-AA produced over time is reproducible.

Example 3-determination of therapeutic concentration in ALS

Patients with ALS receive D2-LA treatment over a period of time. The dose and period of D2-LA administration varies from patient to patient. All patients undergoing evaluation were subjected to ALSFRS-R analysis of their natural course prior to initiation of treatment.

The concentration of D2-AA in red blood cells in blood samples of these patients was assessed. PK sampling of 10 patients showed an average D2-AA concentration in erythrocytes of 3.1% ± 2.0. In addition, the functional score (ALSFRS-R result) of each patient at the end of treatment was compared to the natural course score at the beginning of treatment. Based on this comparison, the rate of decline changed from annual rate of-14.2 +/-4.4 prior to treatment to-7.6 +/-1.4 or 46% decrease during treatment (p=0.07, paired t-test of slope change in subjects).

The results of this study demonstrate that the concentration of D2-AA in erythrocytes is about 3% based on the total amount of arachidonic acid + deuterated arachidonic acid found therein, which is considered therapeutic (including D2-AA), providing a therapeutic effect in reducing the rate of disease progression when the concentration of D2-AA in erythrocytes is at least about 3%.

Example 4-determination of therapeutic concentration in PSP

3 Patients diagnosed with suspected PSP (one example of tauopathies) were baseline assessed using 28 progressive supranuclear palsy scales (PSPRS) [19] and the unified Parkinson's disease scale (UPDRS). Then treated with D2-Lin (2.88 g BID;5.76g total daily dose) and observed for disease progression. During treatment, scores of 2 scoring scales were determined every 3 months. Pharmacokinetic (PK) sampling was performed at month 3. These analytes include plasma and erythrocyte membrane levels (D2-LA) and their central active metabolites D2-AA.

The 3 patients were 2 men (66 years and 73 years) and 1 female (74 years), and the duration of the pre-treatment symptoms between 6 years was 2 men for 3 years and 2 women, respectively. Baseline PSPRS for two men was 17 and 12, respectively, and 13 for women. The baseline UPDRS was 44 and 36 for two men and 21 for women, respectively.

After 3 months of treatment, the slope of PSPRS changed from a historical decline of 0.91 min/month to an average decline of 0.16 min/month (+/-0.23 SEM). The slope of UPDRS changed from the expected 0.95 min/month increase to an average increase of 0.28 min/month (+/-0.41 SEM).

At 3 months, the following data were collected:

* Patient 1 was evaluated at 4 months

At 12 months, the following data were collected:

* Patient 1 was evaluated at 13 months

The second male patient had increased the dose (2.88 g TID;8.64 g total daily dose) after the first year of treatment to further boost his treatment.

The above results demonstrate that a percentage of 13,13-D2-AA to (AA+13, 13-D2) in erythrocytes of about 3% is necessary to achieve a therapeutic effect. This data further demonstrates that a percentage of 13,13-D2-AA to (AA+13, 13-D2-AA) in the erythrocytes of about 5% is preferred; and more preferably 6% or 8%.

Claims

1. A non-invasive method for determining the concentration of deuterated arachidonic acid in reporter cells of a patient population, the concentration being correlated with the occurrence of a therapeutic effect in a patient population having a neurodegenerative disease treatable with said deuterated arachidonic acid, the method comprising:

a) Obtaining data from a population of patients having a neurodegenerative disease for the first time being treated with deuterated arachidonic acid or a prodrug thereof, the data comprising the concentration of deuterated arachidonic acid obtained in a patient-reported cell population one or more times after initiation of treatment;

b) Assessing when a patient exhibits a therapeutic outcome;

e) Repeating steps a) -d) for one or more neurodegenerative diseases;

2. A method for determining a patient's response to administration of a composition comprising deuterated arachidonic acid or a prodrug thereof during treatment of a treatable neurodegenerative disease, the method comprising:

3. The method of claim 1, wherein the sample of reporter cells is obtained about 1 month after initiation of the treatment.

4. The method of claim 3, wherein further samples of the reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

5. The method of any one of claims 1, 3 or 4, wherein the reporter cell is a red blood cell, a skin cell, an adipocyte, a biopsy cell, or an epithelial cell.

6. The method of claim 2 or 5, wherein the deuterated arachidonic acid is 13, 13-D2-arachidonic acid.

7. The method of claim 2 or 5, wherein the deuterated arachidonic acid is D4-arachidonic acid.

8. The method as recited in claim 2 or 5 wherein said deuterated arachidonic acid is D6-arachidonic acid wherein said deuterated D6-arachidonic acid comprises at least 90% of hydrogen atoms replaced with deuterium at position 7,7,10,10,13,13 and optionally up to 35% of the remaining hydrogen atoms at non-bis-allylic positions are replaced with deuterium.

9. A method for assessing a patient's response to administration of a composition comprising deuterated arachidonic acid or a prodrug thereof during treatment of a treatable neurodegenerative disease, the method comprising:

A sub-therapeutic concentration when the concentration of deuterated arachidonic acid in erythrocytes of a patient is lower than the determined therapeutic concentration, and a therapeutic concentration when the concentration of deuterated arachidonic acid is equal to or greater than the determined therapeutic concentration; and

10. The method of claim 9, wherein the sample of reporter cells is obtained about 1 month after initiation of the treatment.

11. The method of claim 10, wherein further samples of the reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

12. The method of any one of claims 9, 10 or 11, wherein the reporter cell is a red blood cell, a skin cell, an adipocyte, a biopsy cell, or an epithelial cell.

13. The method of claim 9, wherein the deuterated arachidonic acid is 13, 13-D2-arachidonic acid.

14. The method of claim 9, wherein the deuterated arachidonic acid is D4-arachidonic acid.

15. The method of claim 9, wherein the deuterated arachidonic acid is D6-arachidonic acid, the deuterated D6-arachidonic acid comprising at least 90% of hydrogen atoms replaced with deuterium at position 7,7,10,10,13,13, and optionally having up to 35% of the remaining hydrogen atoms replaced with deuterium at non-bis-allyl positions.

16. A diagnostic method for determining whether the concentration of deuterated 13, 13-D2-arachidonic acid is therapeutic or sub-therapeutic in a patient having a treatable neurodegenerative disease without intervention in neurons or cerebrospinal fluid of the patient, the method comprising:

Comparing the concentration to a minimum therapeutic concentration of 13, 13-D2-arachidonic acid based on at least 3% of the total amount of arachidonic acid including deuterated arachidonic acid present in the red blood cells to determine whether the patient has a therapeutic or sub-therapeutic concentration of 13, 13-D2-arachidonic acid in the neurons.

17. The method of claim 16, wherein the therapeutic concentration of 13, 13-D2-arachidonic acid is set to 4%, 5%, 6%, 7%, 8%, 9% or even 10% as a therapeutic target for a given patient or group of patients.

18. The method of claim 16, wherein the sample of reporter cells is obtained about 1 month after initiation of the treatment.

19. The method of claim 16, wherein further samples of the reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

20. A diagnostic method for determining whether the concentration of deuterated D4-arachidonic acid in a patient having a treatable neurodegenerative disease is therapeutic or sub-therapeutic without intervention in neurons or cerebrospinal fluid of the patient, the method comprising:

obtaining a blood sample from a patient receiving D4-arachidonic acid or an ester thereof for treatment of a neurodegenerative disease; and

Comparing the concentration to a minimum therapeutic concentration of D4-arachidonic acid based on at least 1% of the total amount of arachidonic acid including deuterated arachidonic acid in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D4-arachidonic acid in neurons.

21. The method of claim 20, wherein the therapeutic concentration of D4-arachidonic acid is set to 2%, 3%, or 5% as a therapeutic target for a given patient or group of patients.

22. The method of claim 20, wherein the sample of reporter cells is obtained about 1 month after initiation of the treatment.

23. The method of claim 20, wherein further samples of the reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

24. A diagnostic method for determining whether the concentration of deuterated D6-arachidonic acid in a patient having a treatable neurodegenerative disease is therapeutic or sub-therapeutic without intervention in neurons or cerebrospinal fluid of the patient, the method comprising:

Comparing the concentration to a minimum therapeutic concentration of D6-arachidonic acid of at least 0.5% based on total amounts of arachidonic acid including deuterated arachidonic acid in the erythrocytes to determine whether the patient has a therapeutic or sub-therapeutic concentration of D6-arachidonic acid in neurons.

25. The method of claim 24, wherein the therapeutic concentration of D6-arachidonic acid is set to 1%, 1.5%, 2%, 2.5%, or 3% as a therapeutic target for a given patient or group of patients.

26. The method of claim 24, wherein the sample of reporter cells is obtained about 1 month after initiation of the treatment.

27. The method of claim 24, wherein further samples of the reporter cells are obtained at intervals of 1 month thereafter, 3 months thereafter, half a year thereafter or each year thereafter.

28. The method of claim 24, wherein the deuterated arachidonic acid is D6-arachidonic acid, wherein the deuterated D6-arachidonic acid comprises at least 90% of hydrogen atoms replaced with deuterium at position 7,7,10,10,13,13, and optionally up to 35% of the remaining hydrogen atoms at non-bis-allyl positions are replaced with deuterium.

29. A kit comprising diagnostic material for performing said method, and a correlation table correlating concentration of deuterated arachidonic acid in erythrocytes to its concentration in human cerebrospinal fluid; in addition to the correlation table, the diagnostic material may include one or more instructions regarding when such a test should be performed, factors suggesting delay of such a test, and the like.