COMPOSITIONS AND METHODS FOR AMELIORATING CNS INFLAMMATION, PSYCHOSIS, DELIRIUM, PTSD or PTSS

• US 20100297070A1
• Filed: 10/18/2008
• Published: 11/25/2010
• Est. Priority Date: 10/19/2007
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Ali, Sameh S., Behrens, Marie Margarita, Dugan, Laura L.
• Granted Patent: US 9,550,827 B2
• Time in Patent Office: Days
• US Class Current: 424/85.2
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 31/12 Ketones
• CPC Class Codes: A61K 31/192 having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid cannabinoids A61K31/658
• CPC Class Codes: A61K 31/194 having two or more carboxyl groups, e.g. succinic, maleic or phthalic acid
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: B01D 21/262 by using a centrifuge
• CPC Class Codes: C07C 2604/00 Fullerenes, e.g. C60 buckminsterfullerene or C70
• CPC Class Codes: C07C 51/47 by solid-liquid treatment; by chemisorption
• CPC Class Codes: C07C 51/487 by treatment giving rise to chemical modification by chemisorption C07C51/47
• CPC Class Codes: C07K 16/248 IL-6
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C12N 5/0619 Neurons
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides compositions and methods for ameliorating, treating, reversing or preventing pathology or inflammation in the central nervous system (CNS), or the brain, caused or mediated by NFkB, IL-6, IL-6-R, NADPH oxidase (Nox), and/or superoxide and/or hydrogen peroxide production by a NADPH oxidase, including for example ameliorating, treating, reversing or preventing schizophrenia, psychosis, delirium, e.g., post-operative delirium, drug-induced psychosis, psychotic features associated with frailty syndrome (FS), aging, depression, dementias; traumatic war neurosis, post traumatic stress disorder (PTSD) or post-traumatic stress syndrome (PTSS), Amyotrophic Lateral Sclerosis (ALS, or Lou Gehrig'"'"'s Disease), and/or Multiple Sclerosis (MS). The invention also provides methods for purifying a C60 fullerene, C3 (tris malonic acid C60) or malonic acid derivatives.

Claims

1. A method for ameliorating, reversing, treating or preventing an inflammation or a central nervous system (CNS) inflammation, a schizophrenia, a psychosis, a delirium, a post-operative delirium, a drug-induced psychosis, a frailty syndrome (FS), psychotic features associated with frailty syndrome (FS), aging, depression, dementias;

trauma, traumatic war neurosis, post traumatic stress disorder (PTSD), post-traumatic stress syndrome (PTSS), Amyotrophic Lateral Sclerosis (ALS, or Lou Gehrig'"'"'s Disease), and/or Multiple Sclerosis (MS), inflammation from CNS infections, and/or cognitive, learning or memory impairments resulting therefrom, in an individual comprising;

(a) (i) providing a composition that

(1) inhibits or decreases the in vivo activity of NFkB, interleukin-6 (IL-6), interleukin-6 receptor (IL-6-R) or a member of the NADPH oxidase enzyme family (Nox), and/or inhibits or decreases superoxide and/or hydrogen peroxide production by a member of the NADPH oxidase enzyme family, or

(2) acts as a superoxide dismutase mimetic to decrease superoxide and/or hydrogen peroxide; and

(ii) administering an effective amount of the composition of (a) to the individual in need thereof;

(b) the method of (a), wherein the individual is a human;

(c) the method of (a) or (b), wherein the composition comprises a pharmaceutical formulation;

(d) the method of (c), wherein the pharmaceutical formulation is formulated for delivery to the brain or a neural cell, or for passing through the blood brain barrier (BBB);

(e) the method of (d), wherein the pharmaceutical formulation is formulated for delivery to a parvalbumin-positive GABA-ergic interneuron;

(f) the method of any of (a) to (c), wherein the pharmaceutical formulation comprises a therapeutic monoclonal antibody specific for and inhibitory to the activity of NFkB. an IL-6 or IL-6-R and/or an NADPH oxidase enzyme;

(g) the method of (f), wherein the therapeutic monoclonal antibody is a humanized or a human antibody;

(h) the method of (f) or (g), wherein the therapeutic monoclonal antibody against the IL-6-R is tocilizumab, or ACTEMRA™

, or the therapeutic monoclonal antibody is against IL-6, or is CNTO-328, a human-mouse chimeric monoclonal antibody (Mab) to IL-6;

(i) the method of (a) or (b), wherein the composition comprises a small molecule;

(j) the method of (i), wherein the small molecule comprises an o-methoxycatechol, an apocynin, a diapocynin, 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), 4-hydroxy-3′

-methoxy-acetophenon, N-Vanillylnonanamide, staurosporine or related compounds;

(k) the method of (a) or (b), wherein the composition comprises an inhibitory nucleic acid molecule to NFkB, IL-6 or NADPH oxidase;

(l) the method of (k), wherein the inhibitory nucleic acid molecule comprises an RNAi molecule, a double-stranded RNA (dsRNA) molecule, an sRNA, a miRNA (microRNA) and/or a short hairpin RNA (shRNA) molecule, or a ribozyme, or a fragment of an NADPH oxidase-encoding nucleic acid;

(m) the method of any of (a) to (l), wherein the member of the NADPH oxidase enzyme family (Nox) is a Nox1, Nox2, Nox3, Nox4 or Nox5 enzyme;

(n) the method of any of (a) to (l), wherein the infection is a viral, bacterial, yeast and/or fungal infection, or a Haemophilus, Cryptococcus, Filobasidiella, Neisseria, Rickettsia or Borrelia infection;

(o) the method of any of (a) to (n), wherein the composition that inhibits or decreases the in vivo activity of NFkB, interleukin-6 (IL-6) or IL-6R is an antibody against NFkB IL-6 or IL-6-R, respectively;

(p) the method of (o), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is interleukin-10 (IL-10);

(q) the method of (p), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is ilodecakin or TENOVIL™

;

or

(r) the method of any of (a) to (n), wherein superoxide dismutase mimetic that decreases superoxide and/or hydrogen peroxide comprises a C60 fullerene, C₃ (tris malonic acid C60) or a malonic acid derivative.

3. A kit comprising

(a) a composition that inhibits NFkB, interleukin-6 (IL-6), interleukin-6 receptor, a member of the NADPH oxidase enzyme family (Nox), and/or superoxide or hydrogen peroxide production by a member of the NADPH oxidase enzyme family (Nox), or a composition that acts as a superoxide dismutase mimetic to decrease superoxide and/or hydrogen peroxide;

(b) the kit of (a) further comprising instructions comprising use of the method of claim 1;

(c) the kit of any of (a) or (b), wherein the member of the NADPH oxidase enzyme family (Nox) is a Nox1, Nox2, Nox3, Nox4 or Nox5 enzyme;

(d) the kit of any of (a) to (c), wherein the composition that inhibits NFkB, interleukin-6 (IL-6), interleukin-6 receptor, a member of the NADPH oxidase enzyme family (Nox), and/or superoxide and/or hydrogen peroxide production by a member of the NADPH oxidase enzyme family (Nox) comprises an antisense nucleic acid, an sRNA, a miRNA or a ribozyme that binds to hybridization to and inhibits or decreases the activity or expression of an antibody the specifically binds to the NFkB, interleukin-6 (IL-6), interleukin-6 receptor, or the member of the NADPH oxidase enzyme family (Nox);

(e) the kit of any of (a) to (c), wherein the composition that inhibits the NFkB, interleukin-6 (IL-6), interleukin-6 receptor, or the member of the NADPH oxidase enzyme family (Nox), and/or superoxide and/or hydrogen peroxide production by a member of the NADPH oxidase enzyme family (Nox) comprises an antibody that specifically binds to the NFkB, interleukin-6 (IL-6), interleukin-6 receptor, or the member of the NADPH oxidase enzyme family (Nox), respectively;

(f) the kit of any of (a) to (e), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is an antibody against IL-6;

(g) the kit of (f), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is IL-10;

(h) the kit of (g), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is ilodecakin or TENOVIL™

;

or

(i) the kit of (a), wherein superoxide dismutase mimetic that decreases superoxide and/or hydrogen peroxide comprises a C60 fullerene, O₃ (tris malonic acid C60) or a malonic acid derivative.

2. A method for protecting the function of, or maintaining the level of activation or activity of cortical inhibitory neurons, or parvalbumin-positive GABA-ergic interneurons, comprising:

(a) (i) providing a composition that

(1) inhibits or decreases the in vivo activity of NFkB, interleukin-6 (IL-6), interleukin-6 receptor (IL-6-R) or a member of the NADPH oxidase enzyme family (Nox), and/or inhibits or decreases superoxide and/or hydrogen peroxide production by a member of the NADPH oxidase enzyme family, or

(2) acts as a superoxide dismutase mimetic to decrease superoxide and/or hydrogen peroxide; and

(ii) contacting the composition of (a) with the cortical inhibitory neuron or parvalbumin-positive GABA-ergic interneuron;

(b) the method of (a), wherein the contacting is in vivo or in vitro;

(c) the method of (b), wherein the contacting is in vivo and the composition of (a) is administered in an effective amount to an individual in need thereof;

(d) the method of (c), wherein the contacting is in vivo to the CNS, or brain cortex, of the individual;

(e) the method of (c) or (d), wherein the individual is a human;

(f) the method of any of (a) to (e), wherein the composition comprises a pharmaceutical formulation;

(g) the method of (f), wherein the pharmaceutical formulation is formulated for delivery to the brain or a neural cell, or for passing through the blood brain barrier (BBB);

(h) the method of (f) or (g), wherein the pharmaceutical formulation is formulated for delivery to a cortical inhibitory neuron or a parvalbumin-positive GABA-ergic interneuron;

(i) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises a therapeutic monoclonal antibody specific for and inhibitory to the activity of NFkB, an IL-6 or IL-6-R or an NADPH oxidase enzyme;

(j) the method of (i), wherein the therapeutic monoclonal antibody is a humanized or a human antibody;

(k) the method of (i) or (j), wherein the therapeutic monoclonal antibody against the IL-6-R is tocilizumab, or ACTEMRA™

, or the therapeutic monoclonal antibody is against IL-6, or is CNTO-328, a human-mouse chimeric monoclonal antibody (Mab) to IL-6;

(l) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises a small molecule;

(m) the method of (l), wherein the small molecule comprises an o-methoxycatechol, an apocynin, a diapocynin, 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), 4-hydroxy-3′

-methoxy-acetophenon, N-Vanillylnonanamide, staurosporine or related compounds;

(n) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises an inhibitory nucleic acid molecule to NFkB, IL-6, IL-6-R, or NADPH oxidase;

(o) the method of (n), wherein the inhibitory nucleic acid molecule comprises an RNAi molecule, a double-stranded RNA (dsRNA) molecule, an sRNA, a miRNA (microRNA) and/or a short hairpin RNA (shRNA) molecule, or a ribozyme, or a fragment of an NADPH oxidase-encoding nucleic acid;

(p) the method of any of (a) to (o), wherein the member of the NADPH oxidase enzyme family (Nox) is a Nox1, Nox2, Nox3, Nox4 or Nox5 enzyme;

(q) the method of any of (a) to (p), wherein the composition that inhibits or decreases the in vivo activity of NFkB, IL-6, IL-6-R, or NADPH oxidase is an antibody against NFkB, IL-6, IL-6-R, or NADPH oxidase, respectively;

(r) the method of (q), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is IL-10;

(s) the method of (r), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is ilodecakin or TENOVIL™

;

or

(t) the method of any of (a) to (h), wherein superoxide dismutase mimetic that decreases superoxide and/or hydrogen peroxide comprises a C60 fullerene, C₃ (tris malonic acid C60) or a malonic acid derivative.

4-7. -7. (canceled)

8. A method for ameliorating (slowing, reversing or abating) or preventing neuron or central nervous system (CNS) or brain damage in individuals having an inflammation or a CNS inflammation, an injury, a pathology, a disease, an infection and/or a condition causing and/or associated with an increased amount of inflammation or CNS inflammation, oxidative stress and/or CNS oxidative stress, or accelerating the recovery of CNS neuron or brain damage in individuals having an inflammation, injuries, pathologies, diseases, infections, and/or cognitive, learning or memory impairments resulting therefrom;

and conditions causing and/or associated with an increased amount of an inflammation, a CNS inflammation and/or CNS oxidative stress, comprising;

(a) (i) providing a composition that inhibits or decreases the level of activation or activity of NFkB, IL-6, IL-6-R, a member of the NADPH oxidase enzyme family (Nox), and/or inhibits or decreases superoxide or hydrogen peroxide production by a member of the NADPH oxidase enzyme family (Nox), or providing a composition that acts as a superoxide dismutase mimetic to decrease superoxide and/or hydrogen peroxide; and

(ii) contacting or administering a therapeutically effective amount of the composition of (a) with an individual in need thereof;

(b) the method of (a), wherein the individual in need thereof is a human;

(c) the method of (a) or (b), wherein the composition of (a) is formulated as a pharmaceutical composition;

(d) the method of any of (a) to (c), wherein the contacting or administering is into the CNS, or brain cortex, of the individual;

(e) the method of any of (a) to (d), wherein the individual is a human;

(f) the method of any of (a) to (e), wherein the human has frailty syndrome (FS), Alzheimer'"'"'s disease, Lewy Body Disease, Parkinson'"'"'s Disease, Huntington'"'"'s Disease, Multi-infarct dementia (vascular dementia), senile dementia or Frontotemporal Dementia (Pick'"'"'s Disease), Amyotrophic Lateral Sclerosis (ALS, or Lou Gehrig'"'"'s Disease), and/or Multiple Sclerosis (MS), and/or cognitive, learning or memory impairments resulting therefrom;

(g) the method of any of (a) to (f), wherein the composition or pharmaceutical formulation is formulated for delivery to the CNS, or brain or a CNS neural cell, or for passing through the blood brain barrier (BBB);

(h) the method of any of (a) to (g), wherein the composition or pharmaceutical formulation is formulated for delivery to a parvalbumin-positive GABA-ergic interneuron;

(i) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises a therapeutic monoclonal antibody specific for and inhibitory to the activity of an NFkB, IL-6 or IL-6-R or an NADPH oxidase (Nox) enzyme;

(j) the method of (i), wherein the therapeutic monoclonal antibody is a humanized or a human antibody;

(k) the method of (i) or (j), wherein the therapeutic monoclonal antibody against the IL-6-R is tocilizumab, or ACTEMRA™

, or the therapeutic monoclonal antibody is against IL-6, or is CNTO-328, a human-mouse chimeric monoclonal antibody (Mab) to IL-6;

(l) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises a small molecule;

(m) the method of (l), wherein the small molecule comprises an o-methoxycatechol, an apocynin, a diapocynin, 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), 4-hydroxy-3′

-methoxy-acetophenon, N-Vanillylnonanamide, staurosporine or related compounds;

(n) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises an inhibitory nucleic acid molecule to the expression of NFkB, IL-6, IL-6-R or NADPH oxidase;

(o) the method of (n), wherein the inhibitory nucleic acid molecule comprises an RNAi molecule, a double-stranded RNA (dsRNA) molecule, an sRNA, a miRNA (microRNA) and/or a short hairpin RNA (shRNA) molecule, or a ribozyme, or a fragment of an NFkB-, IL-6-, IL-6-R- or NADPH oxidase-encoding nucleic acid;

(p) the method of any of (a) to (o), wherein the member of the NADPH oxidase enzyme family (Nox) is a Nox1, Nox2, Nox3, Nox4 or Nox5 enzyme;

(q) the method of any of (a) to (p), wherein the injury is a concussive or traumatic injury, and/or injury is post-concussion syndrome (also known as postconcussive syndrome or PCS), and/or cognitive, learning or memory impairments resulting therefrom;

(r) the method of any of (a) to (p), wherein the infection is a bacterial, viral or yeast infection, or the infection is an HIV infection;

(s) the method of any of (a) to (r), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is an antibody against IL-6;

(t) the method of (s), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is IL-10;

(u) the method of (t), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is ilodecakin or TENOVIL™

;

or

(v) wherein superoxide dismutase mimetic that decreases superoxide and/or hydrogen peroxide comprises a C60 fullerene, C₃ (tris malonic acid C60) or a malonic acid derivative.

9. A method for increasing resistance to or recovery from an inflammation, a CNS injury, a neurological pathology, a disease, an infection and/or a condition caused by and/or associated with an increased amount of inflammation and/or oxidative stress in the CNS or brain, and/or cognitive, learning or memory impairments resulting therefrom, comprising:

(a) (i) providing a composition that inhibits or decreases the level of activation or activity of NFkB, IL-6, IL-6-R, a member of the NADPH oxidase enzyme family (Nox), and/or inhibits or decreases superoxide or hydrogen peroxide production by a member of the NADPH oxidase enzyme family (Nox), or providing a composition that acts as a superoxide dismutase mimetic to decrease superoxide and/or hydrogen peroxide; and

(ii) contacting or administering a therapeutically effective amount of the composition of (a) with an individual in need thereof; and

(ii) contacting or administering a therapeutically effective amount of the composition of (a) with an individual in need thereof;

(b) the method of (a), wherein the individual in need thereof is a human;

(c) the method of (a) or (b), wherein the composition of (a) is formulated as a pharmaceutical composition;

(d) the method of any of (a) to (c), wherein the contacting or administering is into the CNS or brain cortex of the individual;

(e) the method of any of (a) to (d), wherein the individual is a human;

(f) the method of any of (a) to (e), wherein the human has frailty syndrome (FS), Alzheimer'"'"'s disease, Lewy Body Disease, Parkinson'"'"'s Disease, Huntington'"'"'s Disease, Multi-infarct dementia (vascular dementia), senile dementia or Frontotemporal Dementia (Pick'"'"'s Disease), Amyotrophic Lateral Sclerosis (ALS, or Lou Gehrig'"'"'s Disease), and/or Multiple Sclerosis (MS);

(g) the method of any of (a) to (f), wherein the composition or pharmaceutical formulation is formulated for delivery to the CNS or brain or a neural cell, or for passing through the blood brain barrier (BBB);

(h) the method of any of (a) to (g), wherein the composition or pharmaceutical formulation is formulated for delivery to a parvalbumin-positive GABA-ergic interneuron;

(i) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises a therapeutic monoclonal antibody specific for and inhibitory to the activity of an NFkB, IL-6 or IL-6-R or an NADPH oxidase (Nox) enzyme;

(j) the method of (i), wherein the therapeutic monoclonal antibody is a humanized or a human antibody;

(k) the method of (i) or (j), wherein the therapeutic monoclonal antibody against the IL-6-R is tocilizumab, or ACTEMRA™

, or the therapeutic monoclonal antibody is against IL-6, or is CNTO-328, a human-mouse chimeric monoclonal antibody (Mab) to IL-6;

(l) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises a small molecule;

(m) the method of (l), wherein the small molecule comprises an o-methoxycatechol, an apocynin, a diapocynin, 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), 4-hydroxy-3′

-methoxy-acetophenon, N-Vanillylnonanamide, staurosporine or related compounds;

(n) the method of any of (a) to (h), wherein the composition or the pharmaceutical formulation comprises an inhibitory nucleic acid molecule to a nucleic acid encoding an NFkB, IL-6, IL-6-R or NADPH oxidase;

(o) the method of (n), wherein the inhibitory nucleic acid molecule comprises an RNAi molecule, a double-stranded RNA (dsRNA) molecule, an sRNA, an miRNA (microRNA) and/or a short hairpin RNA (shRNA) molecule, or a ribozyme, or an inhibitory fragment of an NFkB-, IL-6-, IL-6-R- or NADPH oxidase-encoding nucleic acid sequence;

(p) the method of any of (a) to (o), wherein the member of the NADPH oxidase enzyme family (Nox) is a Nox1, Nox2, Nox3, Nox4 or Nox5 enzyme;

or

(q) the method of any of (a) to (p), wherein the method increases resistance to a concussive or traumatic injury, and/or the method increases resistance to or ameliorate the effects of post-concussion syndrome (also known as postconcussive syndrome or PCS), and/or cognitive, learning or memory impairments resulting therefrom;

(r) the method of any of (a) to (q), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is an antibody against IL-6;

(s) the method of (r), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is IL-10;

(t) the method of (s), wherein the composition that inhibits or decreases the in vivo activity of interleukin-6 (IL-6) is ilodecakin or TENOVIL™

;

or

(u) the method of (s), wherein the superoxide dismutase mimetic that decreases superoxide and/or hydrogen peroxide comprises a C60 fullerene, C₃ (tris malonic acid C60) or a malonic acid derivative.

10. A method for purifying C60 fullerene derivatives, including C₃ (tris malonic acid C60) and other malonic acid derivatives comprising

(i) (a) dissolving an impure powder form of C₃ (tris malonic acid C60 fullerene) or other malonic acid derivatives in dilute sodium hydroxide (NaOH) solution at a concentration of between about 1 mM to 400 mM at about 4 degrees C. with stirring;

(b) adding a second solution of NaOH more concentrated than the dilute NaOH solution in step (a) drop-wise to the solution of step (a) to achieve an approximately neutral pH;

(c) incubating the solution of step (b) at 4 degrees C. in the dark for approximately 0.5 to 3 hours;

(d) centrifuging the solution after the incubating of step (c) to produce a clear dark red supernatant and a solid light pink pellet;

(e) removing the supernatant to a different container;

(f) incubating the supernatant removed in step (e) at 4 degrees C. for an additional about 3 to 4 hours; and

(g) (1) re-centrifuging to remove substantially all or all undissolved material to generate a pellet and a solution comprising purified C₃, wherein the pellet comprises an insoluble waxy material containing contaminant and small amounts of residual C₃, or (2) filtering the sample through a filter which allows only aqueous solutions to pass, thereby removing an insoluble waxy contaminant after solubilization in dilute NaOH, thereby generating a solution comprising purified C₃;

or

(ii) the method of (i), wherein the purified C₃ solution is further treated to remove a minor amount of volatile contaminant by vacuum distillation or by bubbling an inert gas through the solution.

11. A method for purifying C60 fullerene derivatives, including C₃ (tris malonic acid C60) and other malonic acid derivatives comprising

(a) providing a solution comprising an impure powder form of C₃ (tris malonic acid C60) or other malonic acid derivative;

and

(b) providing an antibody directed against the C60 fullerene derivative, C₃ (tris malonic acid C60) or other malonic acid derivative; and

(c) isolating the C60 fullerene derivative, C₃ (tris malonic acid C60) or other malonic acid derivative by incubating the antibody with the C60 fullerene derivative, C₃ (tris malonic acid C60) or other malonic acid derivative under conditions wherein the antibody specifically binds to the C60 fullerene, to C₃ (tris malonic acid C60) or to another malonic acid derivative;

or

(ii) the method of (i), wherein an antibody-C60 fullerene, antibody-C₃ (tris malonic acid C60) or antibody-malonic acid derivative complex is purified by gel electrophoresis purification, HPLC, immunoprecipitation, column chromatography, differential centrifugation or affinity column chromatography.