WO2015171270A1

WO2015171270A1 - Method for the treatment of neurofibromatosis

Info

Publication number: WO2015171270A1
Application number: PCT/US2015/025645
Authority: WO
Inventors: Mitchell S. Felder
Original assignee: Individual
Current assignee: Individual
Priority date: 2014-05-06
Filing date: 2015-04-14
Publication date: 2015-11-12
Anticipated expiration: 2016-11-06

Abstract

A method of treating neurofibromatoses is presented. The neurofibromatoses are a set of inherited disorders, including neurofibromatosis type 1 (NF1), neurofibromatosis type 2 (NF2), and schwannomatosis, that cause the development of benign tumors of the nerve sheath. Specifically, the treatment method involves the extracorporeal treatment of one or more body fluids in two stages, characterized by removing a body fluid from a living body diseased with a type of neurofibromatosis, passing the body fluid through a first stage, applying an anti-angiogenesis, anti-tumorigenesis, anti-mast cell attractant, and/or activator to at least one antigen in the body fluid, creating an antibody-antigen moiety, passing the treated body fluid through a second stage, removing the antibody-antigen moiety from the body fluid during passage through the second stage, and returning the purified body fluid to the body. The body fluids that can be treated include cerebrospinal fluid (CSF), lymph, and blood.

Description

Method for the Treatment of Neurofibromatosis

Cross-Reference to Related Applications

[001] This application claims benefit under 35 U.S.C. § 119(e) of U.S. Patent Application No. 61/988,941, filed May 6, 2014, which is hereby incorporated herein by reference in its entirety.

Field of the Invention

[002] Generally, the present invention relates to an article and method of sequentially and extracorporeally applying a treatment to a patient's blood and then removing the treatment from the blood before returning the blood to the patient's bloodstream.

Background of the Invention

[003] The neurofibromatoses are a set of inherited disorders, designated as neurofibromatosis type 1 (NFl), neurofibromatosis type 2 (NF2), and schwannomatosis, that classically causes the development of benign tumors of the nerve sheath. Each subtype is distinguished by specific clinical features and is due to mutations in distinct genes.

[004] Neurofibromatosis type 1 is the most common of the disorders, affecting ~1 in 3500 individuals worldwide. The hallmark lesion in NFl is the neurofibroma, whereas schwannomas are characteristic of NF2 and schwannomatosis. These tumors may be differentiated histologically.

[005] Unlike the other two disorders, NFl also includes non-tumor manifestations, making it a true multisystem disorder. Neurofibromatosis (NF, neurofibromatosis type 1) is an inherited disease in which the nerve tissue grows neurofibroma tumors that may be benign, but have the potential for malignant transformation. Neurofibromas may cause severe damage by compressing nerves and other tissues.

[006] Neurofibromatosis is an autosomal dominant disorder. The severity in affected individuals can vary, which is due to variable expressivity.

[007] Approximately half of the cases of newly diagnosed neurofibromatosis are due to de novo mutations and no other affected family members are seen. It affects males and females equally.

[008] Certain molecular organic compounds have been implicated as causing or allowing for the development of neurofibromatosis. There is a continuing need for efficacious treatments for this condition.

Summary of the Invention

[009] The present invention relates to a method of extracorporeal treating a patient's body fluid, including but not limited to, cerebrospinal fluid (CSF), lymph, and blood.

[010] The present invention relates to the treatment of neurofibromatoses, hereinafter abbreviated as "NF." The present invention for the treatment of neurofibromatosis will specifically refer to neurofibromatosis type 1 (NFl), neurofibromatosis type 2 (NF2) and schwannomatosis by name, as necessary.

[011] Specifically, the invention pertains to a method for the extracorporeal treatment of one or more body fluids in two stages characterized by removing a body fluid from a living body diseased with a type of NF, passing the body fluid through a first stage; applying an anti- angiogenesis, anti-tumorigenesis, anti-mast cell attractant and/or activator to at least one, or more, antigen in the body fluid (blood and/or CSF). [012] The treatment comprises creating an antibody-antigen moiety during passage thereof through said first stage; passing the treated body fluid through a second stage; removing antibody-antigen moiety from the body fluid during passage through the second stage, and returning the purified body fluid to the body.

[013] The invention is further characterized by targeting an antigen in the body fluid, with an antibody to allow and facilitate removal thereof in the second stage. The Neurofibromatosis targeted antigen(s) targeted antigens (NF ant.) would include at least one component selected from:

1. Antigens involved in causing or facilitating angiogenesis of

neurofibromas/tumor(s): (NF Ant. Ang.)

A. VEGF: vascular endothelial growth factor

B. Angl: angiopoietin 1

C. Endothelial growth factor

D. Midkine (MK, MDK, neurite growth-promoting factor 2/NEGF 2)

2. Antigens involved in causing or facilitating neurofibromas and/or tumori genesis;

(NF Ant. T.):

A. m-Tor (mammalian target of rapamycin)

B. Fibroblast growth factor-2 (FGF2)

C. Shh protein

D. NGF: nerve growth factor

E. PDGF: platelet-derived growth factor

F. Tumor growth factor, alpha and beta

G. Farnesyltransferase 3. Antigens that are involved in localized mast cell chemoattraction, functioning, and/or activation (NF Ant. Mast):

A. SCF (stem cell factor; Kit-ligand, or KL)

B. Interleukin 1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-17 (IL-17)

C. GROl/CXCLl

D. Tumor necrosis factor-alpha (TNF-a)

4. Antigens that decrease chemotherapeutic efficacy of treatments (NF Ant. Chem.) A. Interleukin-6 (IL-6)

[014] Specifically, the method is further characterized by removing body fluid from a person to produce an extracorporeal bodily fluid; imposing a treatment acting on an antigen of NF in the body fluid, filtering or otherwise removing the treatment from the body fluid, and returning the body fluid to the patient after removing substantially all of the treatment in the second stage.

[015] The method of the present invention comprises treating at least one component of a patient's body fluid extracorporeally with a designer antibody containing an albumin- moiety which will create an albumin-antibody-NF antigen moiety allowing for the efficacious dialysis of the resultant albumin-antibody-NF antigen compound (the targeted NF antigen being

respectively, at least one antigen selected from: NF Ant. Ang., NF Ant. T., NF Ant. Mast, NF Ant. Chem.).

[016] The method of the present invention also allows for the addition of therapeutic compounds during, or in tandem with, the subtractive process. There may be the addition to the body fluid of at least one: 4. Neurofibromin-1 (neurofibromatosis-related protein NF-1) in neurofibromatosis type 1 (NFl)

5. Schwannomin (Merlin; neurofibromin 2) in neurofibromatosis type 2 (NF2)

3. Mast cell inhibitors: cromolyn sodium, and curcumin

4. Cucurbitacin-I

[017] The method of the present invention also allows for the addition of molecular compounds during, or in tandem with, the subtractive process. There may be the addition into the body fluid of:

1. Neurofibromin-1 (neurofibromatosis-related protein NF-1) in neurofibromatosis type 1 (NFl): This may be produced using gene therapy, via an adenovirus, Herpes simplex virus, or retrovirus serving as the vector, thereby enabling the substitution of the NFl gene on the long arm of chromosome 17ql l.2, in neurofibromatosis type 1.

2. Schwannomin (Merlin; neurofibromin 2) in neurofibromatosis type 2 (NF2) : This could be produced using gene therapy, via an adenovirus , Herpes simplex virus, or retrovirus serving as the vector, thereby enabling the substitution of the NF2 gene on chromosome 22ql 1.1, in neurofibromatosis type 2.

[018] More specifically, the method is characterized by removing body fluid from a person to produce an extracorporeal bodily fluid; directing a first antibody against the targeted NF antigen (NF Ant. Ang., NF Ant. T., NF Ant. Mast , NF Ant. Chem.) in the first stage of extracorporeal treatment in the body fluid; in the second stage directing a second antibody conjugated with albumin and/or a protein against the targeted NF antigen thereby forming an albumin-antibody- NF antigen compound; removing at least a substantial portion of the albumin-antibody-NF antigen compound from the body fluid by dialysis, other filtering, or other means; and returning the body fluid to the patient.

[019] The method is characterized by the ability to increase the efficacy of the methodology by using magnetic resonance angiography (MRA) and magnetic resonance venography (MRV) to specifically characterize the arterial and venous flow of the patient's circulation in relation to neurofibromas and/or tumors. This technique may also be used for localized treatment(s), as well as total corporeal treatment.

[020] Also, the method is characterized by testing the blood and/or CSF to determine the efficacy of treatment before returning the body fluid to the patient.

Brief Description of the Drawings

[021] Figure 1 is a partial cross sectional view of a cylinder and tubing used to deliver a treatment to a bodily fluid.

[022] Figure 2 is a partial cross sectional view showing additional detail of the cylinder and tubing of Figure 1.

Detailed Description of the Invention

[023] In the first stage of treatment, a selected body fluid is removed using a standard catheter and/or lumbar puncture. In the second stage, the body fluid is treated with antibodies against the targeted NF antigen (NF Ant. Ang., NF Ant. T., NF Ant. Mast, NF Ant. Chem.).

[024] The method of the present invention comprises treating at least one component of a patient's body fluid extracorporeally with a designer antibody containing an albumin- moiety to create an albumin-antibody-NF antigen moiety allowing for the efficacious dialysis, filtering or other means of removal of the resultant albumin-antibody-NF antigen compound (the targeted NF antigen being respectively, one or a combination of antigen(s) from: NF Ant. Ang., NF Ant. T., NF Ant. Mast , NF Ant. Chem.

[025] The albumin-antibody will be directed towards facilitating removal of the targeted NF antigen(s): (NF Ant. Ang., NF Ant. T., NF Ant. Mast, NF Ant. Chem.). After the removal of the NF antigen(s), the cleansed body fluid will be returned to the patient. The frequency of treatment, and the specifically targeted NF antigen(s) to be removed would depend upon the underlying symptomatology and pathology of the patient, and would be determined by the patient's physician. The article used in performing the method includes two-stages. The first stage includes a treatment chamber for addition of an antibody with an attached albumin moiety, which is added to the body fluid. A second stage receives the treated blood and/or CSF and includes a unit for removing the treatment.

[026] The method includes providing a dialysis or other filtering machine with a first stage and a second stage, and sequentially passing the extracorporeal body fluid through the first and second stages. The body fluid is removed from the patient using standard procedure. The first stage applies a treatment using an antibody which was has attached to it an albumin moiety (or alternatively, a moiety which allows for the efficacious dialysis or removal by other techniques of the antibody-albumin-NF antigen), for the treatment of the body fluid. The second stage substantially removes the treatment. The purified body fluid (body fluid with removed targeted NF antigen: NF Ant. Ang., NF Ant. T., NF Ant. Mast, NF Ant. Chem.) is then tested for the efficacy of removal of the NF antigen and returned to the patient.

[027] An alternative methodology of the present intervention would utilize a designer antibody with an attached macromolecular moiety instead of an albumin moiety. The macromolecular moiety, attached to the antibody, would be 1.000 mm to 0.005 mm in diameter. The antibody- macromolecular moiety-targeted antigen complex would then be blocked from reentering the patient's body fluid circulation, by utilizing a series of microscreens which contain openings with a diameter 50.0000% to 99.9999% less than the diameter of the designer antibody- macromolecular moiety. The microscreen opening(s) must have a diameter of at least 25 microns to allow for the passage and return to circulation of the non-pathology-inducing body fluid constituents.

[028] Alternatively, the target NF antigens may be captured by using antibody microarrays which contain antibodies to targeted NF antigen(s). The antibody microarrays are composed of millions of identical monoclonal antibodies attached at high density on glass or plastic slides. After sufficient extracorporeal exposure of the targeted NF antigens to the antibody microarrays, the antibody microarrays-targeted NF antigens may be disposed of, using standard medical practice.

[029] Another alternative methodology of the present intervention comprises removing one or more of the targeted NF antigens from the body fluid by utilizing a designer antibody containing an iron (Fe) moiety. This will then create a Fe-antibody-antigen complex. This iron containing complex may then be efficaciously removed utilizing a strong, localized magnetic force field.

[030] The device of the invention includes a first stage and a second stage. The first stage applies a treatment of an antibody with an attached albumin moiety targeting the NF antigen(s) specifically exacerbating the pathologic condition. The second stage includes substantial removal of the treatment from the extracorporeal body fluid bodily fluid. As shown in Figure 1 , the first stage can include an exterior wall to define a treatment chamber 5. The treatment conveniently can be applied in the treatment chamber 5. Residence times of the body fluid can be altered by changing the dimensions of the treatment chamber, or by using a dialysis vacuum pump. With reference to Figure 1, body fluid enters the inlet 3, passes through the treatment chamber 5, and exits the outlet 4. In embodiments, the treatment of an antibody with an attached albumin moiety targeting the NF antigen(s) can be applied from a delivery tube 6 located within the treatment chamber 5. An inferior wall 9 defines the delivery tube 6. The delivery tube 6 can include at least one lead 7, 8. The lead 7, 8 can deliver the treatment to the treatment chamber 5. Conveniently, the delivery tubes 6 will have a high contact surface area with the blood and/or CSF. As shown, the delivery tube 6 comprises a helical coil.

[031 ] With reference to Figure 2, when the treatment includes the administration of a designer antibody, the delivery tube 6 can be hollow and the interior wall 9 can define a plurality of holes 21. The designer antibodies can be pumped through the delivery tube 6 to effect a desired concentration of designer anti bodies in the body fluid. The designer antibodies can perfuse through the holes 21. The delivery tube 6 can include any suitable material including, for example, metal, plastic, ceramic or combinations thereof. The delivery tube 6 can also be rigid or flexible. In one embodiment, the delivery tube 6 is a metal tube perforated with a plurality of holes. Alternatively, the delivery tube 6 can be plastic. The antibody with attached albumin moiety, targeting the NF antigen(s) can be delivered in a concurrent or counter-current mode with reference to the body fluid. In counter-current mode, the body fluid enters the treatment chamber 5 at the inlet 3. The designer antibody can enter through a first lead 8 near the outlet 4 of the treatment chamber 5. The blood and/or CSF then passes to the outlet 4 and the designer antibodies pass to the second lead 7 near the inlet 3. The removal module of the second stage substantially removes the designer antibodies-NF antigen molecular compound from the body fluid. [032] The second stage can include a filter, such as a dialysis machine, which is known to one skilled in the art. The second stage can include a molecular filter. For example, molecular adsorbents recirculating system (MARS), which may be compatible and/or synergistic with dialysis equipment. MARS technology can be used to remove small to average sized molecules from the body fluid. Artificial liver filtration presently uses this technique.

[033] The method can include a plurality of steps for removing the targeted NF antigen(s). A first step can include directing a first antibody against the targeted antigen. A second step can include a second antibody. The second antibody can be conjugated with albumin, or alternatively another moiety which allows for efficacious dialysis or filtering of the antibody-NF antigen from the body fluid. The second antibody or antibody-albumen complex combines with the first antibody forming an antibody- antibody-moiety complex. A third step is then used to remove the complex from the blood and/or CSF. This removal is enabled by using dialysis and/or MARS. The purified body fluid is then returned to the patient.

[034] In practice, a portion of the purified body fluid can be tested to ensure a sufficient portion of the targeted NF antigen(s) have been successfully removed from the body fluid. Testing can determine the length of treatment and evaluate the efficacy of the sequential dialysis

methodology in removing the targeted NF antigen(s) and suggest the need for further treatment. Body fluid with an unacceptably large concentration of NF complex remaining can then be retreated and refiltered before returning the body fluid to the patient.

[035] In embodiments, the second stage to remove the antibody-moiety-targeted NF antigen complex from the body fluid can be accomplished by various techniques including, for example, dialysis, filtering based on molecular size, protein binding, solubility, chemical reactivity, and combinations thereof. For example, a filter can include a molecular sieve, such as zeolite, or porous membranes that capture complexes comprising molecules above a certain size.

Membranes can comprise polyacrylonitrile, polysulfone, polyamides, cellulose, cellulose acetate, polyacrylates, polymethylmethacrylates, and combinations thereof. Increasing the flow rate or diasylate flow rate can increase the rate of removal of the antibody with attached albumin moiety targeting the NF antigen(s) such as NF Ant. Ang., NF Ant. T., NF Ant. Mast , NF Ant. Chem.).

[036] Further techniques may include continuous renal replacement therapy (CRRT) which can remove large quantities of filterable molecules from the extracorporeal body fluid. CRRT would be particularly useful for molecular compounds that are not strongly bound to plasma proteins. Categories of CRRT include continuous arteriovenous hemofiltration, continuous venovenous hemofiltration, continuous arteriovenous hemodiafiltration, slow continuous filtration, continuous arteriovenous high-flux hemodialysis, and continuous venovenous high flux hemodialysis. The sieving coefficient (SC) is the ratio of the molecular concentration in the filtrate to the incoming body fluid. A SC close to zero implies that the moiety-antibody-targeted antigen complex will not pass through the filter. A filtration rate of 50 ml per minute is generally satisfactory. Other methods of increasing the removability of the antibody-targeted antigen moiety include the use of temporary acidification of the body fluid extracorporeally using organic acids to compete with protein binding sites.

[037] Embodiments of the present invention include the following:

[038] A method for the extracorporeal treatment of a body fluid comprising:

a. applying a treatment to at least one NF antigen in the body fluid to create an antibody- NF antigen moiety; and b. substantially removing the antibody-NF antigen moiety from the body fluid.

[039] A method for the extracorporeal treatment of a body fluid comprising:

a. applying a treatment to at least one NF antigen in the body fluid to create an antibody- NF antigen moiety; and

b. substantially removing the antibody-NF antigen moiety from the body fluid, c. wherein the NF antigen is selected from a group consisting of angiogenesis (NF Ant.

Ang.), tumorigenesis (NF Ant. T.), localized mast cell chemoattraction, (NF Ant. Mast), decreased chemotherapeutic efficacy (NF Ant. Chem.), and combinations thereof.

[040] A method for the extracorporeal treatment of a body fluid comprising:

a. directing a first antibody against at least one targeted NF antigen;

b. directing a second antibody conjugated with a protein against the targeted antigen, thereby forming an protein-antibody-NF antigen compound; and

c. removing at least a portion of the protein-antibody-NF antigen compound from the body fluid.

[041] A method for the extracorporeal treatment of a body fluid comprising:

a. directing a first antibody against at least one targeted NF antigen;

b. directing a second antibody conjugated with a protein against the targeted antigen, thereby forming an protein-antibody-NF antigen compound; and c. removing at least a portion of the protein-antibody-NF antigen compound from the body fluid,

d. wherein the protein comprises albumin.

[042] A method for the extracorporeal treatment of a body fluid comprising:

b. substantially removing the antibody-NF antigen moiety from the body fluid

c. wherein the body fluid is removed from a patient before applying the treatment and the body fluid is returned to the patient after substantially removing the antibody-NF antigen moiety from the body fluid.

[043] A method for the extracorporeal treatment of a body fluid comprising:

b. substantially removing the antibody-NF antigen moiety from the body fluid

c. wherein the body fluid is removed from a patient before applying the treatment and the body fluid is returned to the patient after substantially removing the antibody-NF antigen moiety from the body fluid,

d. characterized by testing the body fluid for the efficacy of treatment before returning the body fluid to the patient. [044] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about."

[045] Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention.

[046] At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[047] While the invention has been particularly shown and described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention. All documents, books, manuals, papers, patents, published patent applications, guides, abstracts, and other references cited herein are incorporated by reference in their entirety.

[048] Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with the true scope and spirit of the invention being indicated by the following claims.

Claims

Claims:

1. A method for the extracorporeal treatment of a body fluid comprising:

a. applying a treatment to at least one NF antigen in the body fluid to create an

antibody-NF antigen moiety; and

b. substantially removing the antibody-NF antigen moiety from the body fluid.

2. The method of claim 1 wherein the NF antigen is selected from a group consisting of

angiogenesis (NF Ant. Ang.), tumorigenesis (NF Ant. T.), localized mast cell

chemoattraction, (NF Ant. Mast), decrease chemotherapeutic efficacy (NF Ant. Chem.), and combinations thereof.

3. A method for the extracorporeal treatment of a body fluid comprising:

a. directing a first antibody against at least one targeted NF antigen;

4. The method of claim 3, wherein the protein comprises albumin. The method of claim 1 , wherein the body fluid is removed from a patient before applying the treatment and the body fluid is returned to the patient after substantially removing the antibody-NF antigen moiety from the body fluid.

The method of claim 5 characterized by testing the body fluid for the efficacy of treatment before returning the body fluid to the patient.