WO2016118956A1 - Effecteurs pharmaceutiques à modulation idéotypique pour traitement cellulaire sélectif - Google Patents

Effecteurs pharmaceutiques à modulation idéotypique pour traitement cellulaire sélectif Download PDF

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Publication number
WO2016118956A1
WO2016118956A1 PCT/US2016/014751 US2016014751W WO2016118956A1 WO 2016118956 A1 WO2016118956 A1 WO 2016118956A1 US 2016014751 W US2016014751 W US 2016014751W WO 2016118956 A1 WO2016118956 A1 WO 2016118956A1
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cells
imps
domain
detection
imp
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PCT/US2016/014751
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English (en)
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Jacob Jennings ORME
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Imperium Biotechnologies, Inc.
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Priority claimed from US14/603,691 external-priority patent/US20150290284A1/en
Application filed by Imperium Biotechnologies, Inc. filed Critical Imperium Biotechnologies, Inc.
Publication of WO2016118956A1 publication Critical patent/WO2016118956A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4705—Regulators; Modulating activity stimulating, promoting or activating activity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention is related generally to ideotype-specific treatments of cells and organisms, and more particularly to ideotype-specific treatments of cells and organisms using engineered Ideotypically Modulated Pharmacoeffectors (IMPs).
  • IMPs Ideotypically Modulated Pharmacoeffectors
  • each cell type expresses a unique assortment of proteins, lipids, sugars, nucleotide sequences, and other metabolites. Each of these is a potential antigen, having epitopes with which a molecule having predetermined affinity can interact.
  • the expression of said antigens is modified by the status of the cell and by its environment. This expression becomes further modified when viruses or intracellular bacteria introduce foreign materials into the cell as they infect. Viruses in particular hijack the cell machinery and produce many virion copies that bud off from the cell and infect other cells.
  • a method includes introducing a plurality of engineered Ideotypically Modulated Pharmacoeffectors (IMPs) into a population of cells.
  • IMPs Ideotypically Modulated Pharmacoeffectors
  • Each IMP may include a detection domain and an activation domain.
  • One or more epitopes is bound by the detection domain.
  • the activation domain is activated in response to the binding.
  • each of a plurality of Ideotypically Modulated Pharmacoeffectors include a detection domain and an activation domain.
  • the detection domain has affinity for an epitope.
  • the activation domain is configured such when it is activated, it will cause a downstream effect in a population of cells.
  • each of the plurality of Ideotypically Modulated Pharmacoeffectors includes a detection domain and an activation domain.
  • the detection domain of each IMP of a subset of the plurality of manufactured IMPs is configured to bind to one or more epitopes.
  • the activation domain of each IMP of the subset of the plurality of manufactured IMPs is configured to activate and cause a downstream effect in a population of cells.
  • Another aspect provides a method comprising: introducing a plurality of subunits to a membrane, wherein at least one subunit of the plurality of subunits localizes to the membrane; and, activating at least one localized subunit of the plurality of subunits, so that the at least one localized subunit forms a structure at said membrane when activated to do so.
  • Still another aspect provides a method comprising: introducing a plurality of engineered Ideotypically Modulated Pharmacoeffectors (IMPs) into a population of cells, each IMP including: at least one detection domain having an affinity for one or more epitopes, and at least one porin activation domain coupled to the at least one detection domain; binding at least one detection domain of at least one IMP of a subset of the plurality of IMPs to at least one epitope; and, in response to binding the at least one detection domain, activating the at least one porin subunit activation domain of at least one IMP of the subset of the plurality of IMPs, the activating causing a downstream effect in at least a subset of the population of cells.
  • IMPs engineered Ideotypically Modulated Pharmacoeffectors
  • a further aspect provides a method comprising: introducing a plurality of engineered Ideotypically Modulated Pharmacoeffectors (IMPs) into a population of cells, each IMP including: at least one detection domain having affinity for an epitope, wherein the at least one detection domain binds single-stranded nucleotide sequences, and at least one activation domain coupled to the at least one detection domain; binding at least one detection domain of at least one IMP of a subset of the plurality of IMPs to at least one epitope; and, in response to binding the at least one detection domain, activating the at least one activation domain of at least one IMP of the subset of the plurality of IMPs, the activating causing a downstream effect in at least a subset of the population of cells.
  • IMPs engineered Ideotypically Modulated Pharmacoeffectors
  • Still a further aspect provides a method comprising: producing a plurality of subunits, wherein: at least one subunit of the plurality of subunits is configured to localize to a membrane, at least one subunit of the plurality of subunits is modified to prevent spontaneous oligomerization, and at least one subunit of the plurality of subunits is modified to include at least one detection domain; binding at least one detection domain of at least one of the plurality of subunits to at least one epitope; and, in response to binding the at least one detection domain, oligomerization of adjacent subunits to form a structure.
  • Another aspect provides a method comprising: producing a multiple- IMP pore, wherein: at least one IMP of a plurality of IMPs localizes to a membrane, at least one IMP of a plurality of IMPs oligomerizes, and at least one IMP of a plurality of IMPs forms a pore structure.
  • Certain embodiments of the method may have a number of technical advantages. For example, some embodiments may be capable of terminating diseased or disease-causing cells. Some other embodiments may include enhancing cells. Some further embodiments may be capable of eliminating carriers of zoonotic diseases. Still other embodiments may reduce complications associated with transplants. Various embodiments may include some, all, or none of the above advantages. Particular embodiments may include other advantages.
  • Figure 1 shows one example of an engineered Ideotypically Modulated Pharmacoeffector including a detection domain and an activation domain according to one embodiment
  • Figure 2 A shows one example of multiple unbound Ideotypically Modulated Pharmacoeffectors that may be introduced into a population of cells according to one embodiment
  • Figure 2B shows one example of binding of multiple adjacent ideotypical epitopes to the detection domains of multiple Ideotypically Modulated Pharmacoeffectors according to one embodiment
  • Figure 2C shows the interaction of multimerized Ideotypically Modulated Pharmacoeffectors with endogenous cascade mediators according to one embodiment
  • Figure3A shows one example of multiple unbound Ideotypically Modulated Pharmacoeffectors that may be introduced into a population of cells according to one embodiment
  • Figure 3B shows one example of binding of multiple adjacent ideotypical epitopes to the detection domains of multiple Ideotypically Modulated Pharmacoeffectors according to one embodiment
  • Figure 3C shows the interaction of multimerized Ideotypically Modulated Pharmacoeffectors with endogenous cascade mediators according to one embodiment
  • Figure 4 shows an antibody or antibody-like complex capable of blocking the pore of a polymer pouch containing a number of effector molecules according to one embodiment
  • Figure 5 shows a detection domain linked to an activation domain by a linker according to one embodiment
  • Figure 6 shows the binding of a plurality of Ideotypically Modulated Pharmacoeffectors to the surface antigen receptors of ideotypical disease-causing B cells according to one embodiment
  • Figure 7 shows an example plasmid construct of Ideotypically Modulated Pharmacoeffectors, giving instructions to target cells for the introduction of Ideotypically Modulated Pharmacoeffectors into the cell according to one embodiment
  • Figure 8A shows an example of the products of an example plasmid construct from Figure 7.
  • Figure 8B shows an example of a self-assembled combination of parts shown in Figure 8A.
  • Figure 9 shows one example of a membrane separating two spaces according to one embodiment.
  • Figure 10A shows one example of an engineered Ideotypically Modulated Pharmacoeffector including a detection domain and an activation domain that is configured for introduction into a population of cells according to one embodiment;
  • Figure 10B shows one example of an antigen with one or more epitopes according to one embodiment
  • Figure I OC shows one example of binding of multiple adjacent ideotypical epitopes to detection domains of multiple Ideotypically Modulated Pharmacoeffectors according to one embodiment
  • Figure 10D shows one example of binding of multiple adjacent ideotypical epitopes to detection domains of multiple Ideotypically Modulated Pharmacoeffectors to form a structure according to one embodiment
  • Figure 1 1A shows one example of an engineered Ideotypically Modulated Pharmacoeffector including a detection domain and an activation domain that is configured for introduction into a population of cells according to one embodiment
  • Figure 1 IB shows one example of an antigen with one or more epitopes according to one embodiment
  • Figure 1 1C shows one example of binding of multiple adjacent ideotypical epitopes to detection domains of multiple Ideotypically Modulated Pharmacoeffectors according to one embodiment
  • Figure 1 1D shows one example of binding of multiple adjacent ideotypical epitopes to detection domains of multiple Ideotypically Modulated Pharmacoeffectors to form a structure according to one embodiment
  • Figure 12A shows one example of an engineered Ideotypically Modulated Pharmacoeffector including a detection domain and an activation domain that has been introduced to at least one cell of a population of cells according to one embodiment
  • Figure 12B shows one example of a cell containing an antigen with one or more epitopes, and one or more Ideotypically Modulated Pharmacoeffectors according to one embodiment
  • Figure 12C shows one example of binding of multiple adjacent ideotypical epitopes to detection domains of multiple Ideotypically Modulated Pharmacoeffectors to form a structure in and/or on a membrane according to one embodiment
  • Figure 12D shows one example of binding of multiple adjacent ideotypical epitopes to detection domains of multiple Ideotypically Modulated Pharmacoeffectors to form a structure in and/or on a membrane according to one embodiment
  • Figure 13 A shows one example of part of a model porin monomer according to one embodiment.
  • Figure 13B shows the natural oligomerization of a multiplicity of porin monomers in the lipid bilayer of a cell.
  • Figure 13C shows an Ideotypically Modulated Pharmacoeffector that includes an activation domain comprising part or all of a porin with modification of amino acids involved in oligomerization and/or functionalization for the attachment of one or more detection domains according to one embodiment.
  • Figure 13D shows Ideotypically Modulated Pharmacoeffectors comprising some or all of the modifications in Figure 13C that have oligomerized or oligomerized more readily as a result of the binding of detection domains to antigens with epitopes found in a cell according to one embodiment.
  • FIGURES 1 through 13D discussed below, and the various embodiments used to describe the principles of the present disclosure in this patent document are by way of illustration only and should not be construed in any way to limit the scope of the disclosure. Those skilled in the art will understand that the principles of the present disclosure may be implemented in any suitably arranged configuration. The invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, the disclosed embodiments are provided such that this disclosure will be thorough and complete, and will convey the scope of the invention to those skilled in the art. The principles and features of the invention may be employed in varied and numerous embodiments without departing from the scope of the invention.
  • FIG. 1 shows one example of an engineered Ideotypically Modulated Pharmacoeffector (1) according to one embodiment.
  • an IMP (1) includes a detection domain (2) and an activation domain (3).
  • detection domain refers to any molecule (e.g. protein, nucleotide sequence, lipid) that has affinity for a molecule of interest.
  • activation domain refers to any molecule (e.g. protein, nucleotide sequence, substance) that may be activated to interact with the cell or its environment in response to the binding of the detection domain.
  • "engineered” Ideotypically Modulated Pharmacoeffectors or IMPs (1) may refer to a nonnaturally occurring manufacture or composition of biological matter having a distinctive use.
  • IMPs or IMP -producing substances e.g. nucleotides
  • a plasmid encoding IMPs may be introduced.
  • Figure 2A shows one example of multiple unbound Ideotypically Modulated Pharmacoeffectors (la, lb) that may be introduced into a population of cells according to one embodiment.
  • the detection domains (2a, 2b) of these IMPs (la, lb) have affinity for adjacent epitopes on a predetermined target antigen (4) when present in the cell of a particular ideotype.
  • the term "epitope” as used herein refers to any part of a molecule of interest with which a detection domain, as defined above, interacts or binds.
  • the terms “ideotype,” “ideotypical,” and “ideotypically” as used herein refer to the uniqueness or differential expression of the set of antigens (and their concomitant epitopes) expressed in or on a subset of cells.
  • the term “antigen” as used herein refers to any molecule of interest with epitopes.
  • the detection domains (2a,2b) may be identical or different, depending on the application. For instance, in detecting antigens (4) with repetitive epitopes, identical detection domains (2a,2b) may be advantageous.
  • activation domains 3a,3b
  • the terms "dimerization” and multimerization” as used herein refer to the colocalization of molecules, whether homologous or heterologous, changing the activity of said molecules.
  • activation domains may be identical or different, depending on the application. For instance, an activation domain (3a,3b) for inducing cell death may be inactive Caspase- 9 monomer, which— when dimerized— converts itself to an active form. In this case, an identical activation domain may be advantageous.
  • FIG. 2C shows the interaction of multimerized Ideotypically Modulated Pharmacoeffectors with endogenous cascade mediators (5) from a cell according to one embodiment.
  • This induces a cascade of desired downstream effects in cells which contain the epitopes of the antigen of interest (4).
  • the term "downstream effects" as used herein may refer to any biological result of the interaction of activation domains (3a,3b) with endogenous molecules (5).
  • the epitopes of the antigen of interest (4) may be expressed in a desired target cell type (i.e. ideotype), whether viral, cancerous, or otherwise of interest.
  • a linker of some sort e.g. collagen
  • the length of this linker may be adjusted to maximize downstream effect.
  • Ideotypically Modulated Pharmacoeffectors (1) may be manufactured.
  • a FAb antibody fragment with specific affinity for the target epitopes may be covalently conjugated to an effector domain by a collagen linker by synthetic conjugating processes.
  • an aminophosphonate group may chemically connect the C terminus of a FAb fragment with the N terminus of an inactive Caspase.
  • the genetic code for the entire complex may be introduced in a bacterial, human, or other organic species that may mass-produce the complex as a unit or as subunits to be conjugated later.
  • an anti-sense nucleotide sequence specific for target epitopes of nucleotide sequences (2a, 2b) in the cell could be chemically conjugated to an activation domain (3a, 3b). This may be accomplished through carbodiimides or other available means of conjugation.
  • a nucleotide detection domain (2a,2b) contains an azide that may be linked via Staudinger ligation chemistry to activated ester groups on an activation domain (3a,3b).
  • a nucleotide detection domain (2a, 2b) may contain a benzaldehyde group that may be linked via modified Pictet-Spengler ligation in glacial acetic acid to terminal histidine residues on an activation domain (3a,3b).
  • Both of the aforementioned ligation methods have been used to produce working prototypes by the applicant of the present disclosure.
  • other ligation methods may be utilized. These instances are only some of the ways in which the embodiment could be manufactured and should not be considered limiting. Further examples of manufacturing of IMPs are described below with reference to Figures 2A-6.
  • Figure 3A shows one example of multiple unbound Ideotypically Modulated Pharmacoeffectors (la, lb) that may be introduced into a population of cells according to one embodiment.
  • detection domains (2a,2b) may comprise antisense nucleotide strands.
  • antisense and “complementary” as used herein refer to nucleotide sequences of any length with coded affinity for another nucleotide strand; as a non-limiting example, the sequence 5'-ATGCC-3' could be complementary or antisense to the sequence 5'-GGCAT-3'.
  • activation domains (3a,3b) may perform substantially the same functions as the activation domain shown in Figures 1-2C. These elements may be conjugated (2a,3a and 2b,3b) to form an IMP (la,lb) by carbodiimides or by some other functional group or linker in a chemical process.
  • FIG. 3B shows one example of binding of multiple adjacent ideotypical epitopes (4) to the detection domains (2a,2b) of multiple Ideotypically Modulated Pharmacoeffectors according to one embodiment.
  • epitopes (4) may comprise nucleotide strands that may be unique to target cells of interest.
  • these strands may comprise a nucleotide sequence introduced by a virus as it infects a cell.
  • the detection domains (2a,2b) may be antisense nucleotide sequences that are complementary to that introduced by the virus.
  • Figure 3C shows the interaction of multimerized Ideotypically Modulated Pharmacoeffectors (la, lb) with endogenous cascade mediators (5) according to one embodiment.
  • the nucleotide epitope (4) may be expressed in a particular target cell type (i.e. ideotype), whether viral, cancerous, or otherwise of interest.
  • the nucleotide epitope may be expressed in varying levels in a particular target cell type or ideotype.
  • Figure 4 shows an antibody or antibody -like complex capable of blocking the pore (7) of a polymer pouch (6) containing a number of effector molecules (9) according to one embodiment.
  • these antibodies or antibody-like complexes are held in the pores (7) by Fc-associated holder molecules (8).
  • this holder molecule may be Surface Protein A of Staphylococcus aureus. This blockage of the pore (7) may keep the effector molecules (8) from interacting with the cellular environment.
  • the detection domain (2) of an embodiment may be specific for an epitope of interest, as those mentioned in previous embodiments. Binding of an epitope of interest induces conformational changes in the remainder of the complex, the activation domain (3).
  • Figure 7 shows an example plasmid construct of Ideotypically Modulated Pharmacoeffectors (13), giving instructions to target cells for the introduction of Ideotypically Modulated Pharmacoeffectors into cells according to one embodiment.
  • Various nucleotide sequences found on a plasmid (15) may, among other functions, promote the expression of nucleotide sequence detection domains (2a, 2b) and a protein activation domain (3).
  • an activation domain (3a,3b) may comprise a fusion of a ribonucleoprotein subunit (17a,b) and a caspase.
  • ribonucleoprotein and “nucleoprotein” as used herein refer to any nucleotide-binding protein or protein subunit; they may be produced artificially or are found naturally (e.g. riboswitches). Such a fusion can be produced using widely-available recombinant gene techniques.
  • a detection domain (2a,2b) may comprise a fusion of a ribonucleoprotein-binding specific ribonucleotide sequence (16a, 16b) and a sequence that is complementary to a target sequence found in cells of interest.
  • a cell may be induced by a plasmid to introduce both detection and activation domains into the cell.
  • an IMP may self-assemble as shown in Figure 8B and function much like an embodiment described in Figure 3C.
  • Various uptake sequences (14) may be part of the plasmid to induce uptake into the cell.
  • uptake sequence refers to sequences by which the cell is induced to import the plasmid into the cell.
  • Modified viral vectors may offer cell-specific or generalized introduction of the embodiments into the cell.
  • Such constructs have been developed for gene therapy and may be modified for this purpose. Rather than injecting nucleotide material to be inserted into the host genome, these constructs may contain IMPs.
  • Other potential methods include receptor-mediated endocytosis, which is also used by cells and viruses alike to take in exogenous materials.
  • an IMP may have a subunit with affinity for a surface receptor on all or a subset of cells. As a result of binding, the IMP may be brought into the cell.
  • a plasmid may contain a sequence (“uptake sequence") that is recognized and taken up by some cells.
  • the term "uptake sequence” as used herein refers to sequences by which the cell is induced to import the plasmid into the cell.
  • an embodiment may be encapsulated in a colloid suspension, which may make the embodiment sufficiently amphiphilic to allow association with the lipid bilayer.
  • liposomes containing an IMP or IMPs may be coated with materials that may make them more likely to be taken up by a cell.
  • an array of positive charges on a liposome containing an embodiment may allow non-specific cell bilayer association and fusion. The association of particular peptides may also help to encourage uptake of liposome contents. Different cellular compartments may require different delivery methods.
  • Embodiments as may be used in viral diseases are provided.
  • viral diseases including but not limited to HIV may be treated.
  • the use of an embodiment like those shown in Figures 1 and 2 may cause HIV-infected cells to die by apoptosis.
  • apoptosis refers to programmed cell death in which the cell is induced by internal and/or extemal signaling to die.
  • apoptose refers to the action of apoptosis of a cell.
  • the detection domain (2) of the Ideotypically Modulated Pharmacoeffectors (1) could be antibody fragments with specificity for adjacent epitopes (4) on HIV Reverse Transcriptase (HIV RT).
  • HIV RT expresses epitopes that are unique and may not be found in normal, uninfected human cells. This may allow the differentiation of two ideotypes of cells: infected and uninfected. As previously described, the term "ideotype" as used herein refers to the uniqueness of the set of antigens expressed in or on a subset of cells. HIV RT is necessary for the virus to mount a successful cell invasion and will therefore be found in every HIV-infected cell.
  • the activation domain (3) of an embodiment may be Fas-associated protein with death domain (FADD). When FADD trimerizes naturally in a cell in sufficient numbers, it causes the assembly of the death inducing signaling complexes (DISCs) that signal downstream to terminate the cell.
  • FADD Fas-associated protein with death domain
  • the three FADD activation domains (3) may trimerize to cause downstream DISC formation. This may cause apoptosis of those cells in which sufficient DISCs are formed, which may be cells of the infected ideotype.
  • Detection domains (2a,2b) may comprise antisense nucleotide strands of RNA or DNA as shown in Figures 3A-C.
  • a detection domain (2a,2b) of an embodiment may comprise antisense nucleotide strands that are complementary to adjacent conserved sequences of ideotypical viral nucleotides (4). These strands could be manufactured by using PCR or by transfecting bacteria to produce them.
  • the activation domain (3a,3b) of an embodiment may be an inactive monomer of Caspase-9.
  • a detection domain (2) and an activation domain (3a,3b) may be conjugated together through a benzaldehyde group interacting to an N-terminal histamine residue on the activation domain using a modified Pictet-Spengler reaction in glacial acetic acid.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of an HIV-specific nucleotide strand (4) may bring the activation domains (3a,3b) into contact such that they may dimerize. The cell may apoptose when this occurs in sufficient numbers.
  • Detection domains (2a,2b) may comprise antisense nucleotide strands of RNA or DNA as shown in Figures 3A-C.
  • a detection domain (2) and an activation domain (3a,3b) may be encoded for on a plasmid, as in one embodiment shown in Figure 7.
  • the plasmid may contain a sequence that enhances uptake into cells of interest.
  • the plasmid may encode detection (2a,2b) and activation (3) domains as shown in Figure 8A.
  • an activation domain may comprise a fusion of a procaspase-9 subunit and a ribonucleoprotein subunit (17a,b).
  • a detection domain may comprise a fusion of anti- sense nucleotide strands that recognize an HIV-specific nucleotide strand and a ribonucleoprotein-binding strand (16a,b) that is specific for the ribonucleoprotein subunit found in an activation domain.
  • a cell containing a plasmid produces the domains, which self-assemble as shown in Figure 8B by the interaction of a ribonucleoprotein-binding nucleotide portion (16a,b) of a detection domain with a ribonucleoprotein subunit portion (17a,b) of an activation domain.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of an HIV-specific nucleotide strand (4) may bring the activation domains (3a,3b) into contact such that they may dimerize. The cell may apoptose when this occurs in sufficient numbers.
  • IMPs Ideotypically Modulated Pharmacoeffectors
  • various embodiments of the present disclosure may allow the termination of infected cells before escape variants can be produced by the virus. Since the epitopes detected by IMPs may be selected such that they are conserved, even widely-mutated strains of any given virus may still express them. Further, all of the viral strains in a given cell— even those which have mutations that might make them superior escape variants— may be destroyed when a cell apoptoses. This means that IMPs, unlike existing treatments, may not naturally select evolutionarily superior strains of a virus.
  • IMPs may not cause nor be affected by resistance mechanisms developed by the virus. This is in direct contrast to presently-available antivirals and antibiotics.
  • Another advantage of IMPs may be their scope of effectiveness. While most current antiviral medications only interfere with the growth of the virus, IMPs may clear an infection completely. This gives the further advantage that IMPs may be much more cost-effective than existing treatments, since IMPs may not necessitate long-term treatment. In addition, the complete clearance of disease may eliminate the risk for further transmission.
  • IMPs may avoid the unpleasant side effects of other treatments. These may include lipodystrophy, liver toxicity, and inflammation, amongst others. These side-effects lead a large fraction of patients to forego treatment (e.g. 25% of patients undergoing HAART for HIV infections). Without these side-effects, IMPs may result in higher treatment compliance.
  • IMPs that would target HIV epitopes
  • these and other embodiments may be adjusted for other viral diseases.
  • This could include, for example, viruses that cause the common cold, Dengue fever, Epstein- Barr, HPV, Hantavirus, various forms of hepatitis, herpes, influenza, rotavirus, and others.
  • viruses that cause the common cold, Dengue fever, Epstein- Barr, HPV, Hantavirus, various forms of hepatitis, herpes, influenza, rotavirus, and others.
  • Various embodiments may also be used simultaneously (i.e. various detection domains and a variety of activation domains). This may give an advantage in that it may provide redundancy against potential resistance mechanisms.
  • IMPs could also work against intracellular disease-causing organisms like bacteria or parasites. These disease-causing pathogens, including Rickettsiae and Chlamydia species, can pose problems similar to those of viruses. They also express unique epitopes that may not be found in normal, uninfected cells. As previously, cells can be categorized into ideotypes of infected and uninfected.
  • a detection domain (2) that is a lectin.
  • Lectins are found in nature and are capable of binding specific sugars, as are frequently expressed on the surface of specific bacteria. Some lectins, for instance, are quite specific to the sugars expressed on species of Chlamydia.
  • the activation domain (3) in this embodiment may include various antimicrobials.
  • the effector molecules (9) may include antimicrobials like peptidoglycan lyase.
  • Peptidoglycan lyase is an enzyme that digests the protective barrier found on the surface of most bacteria. On binding of lectins (2) to the specific sugars found on an invading bacterial cell wall in ideotypical cells, peptidoglycan lyase would be freed to digest bacterial cell barriers.
  • IMPs could also be used to treat and/or prevent cancers.
  • Malignant cells may be ideotypical in that they may express epitopes that are either unique or enriched to these cells.
  • the term "malignant cell” or "cancerous cell” as used herein refers to any cell that proliferates out of control as a result of genetic and/or metabolic changes in the cell.
  • These epitopes may vary in their expression levels and distribution depending on the situation. Some epitopes are expressed in all or most cancer types, while others are only present in few. Some epitopes may never be expressed in non-malignant cells, while others may only have elevated expression levels in malignant cells.
  • a detection domain (2) that is an antibody fragment specific for a mutated form of the protein p53 (4) found in some cancers.
  • An activation domain (3) may comprise inactive Caspase-9 monomer.
  • the detection domains (2) of multiple IMPs (1) may bind the mutated p53 (4).
  • Their activation domains (3) may dimerize to cause downstream signaling in the cell. When this signaling reaches sufficient levels, the cell may apoptose.
  • a detection domain (2) may comprise an antibody fragment specific for PSA (prostate specific antigen). While PSA is ideotypical of all prostate cells rather than just cancerous cells, it may be more effective in coping with prostate cancer after it has been diagnosed.
  • An activation domain (3) may comprise inactive Caspase-9 monomer. If administered in smaller amounts, the embodiment may lower the threshold of natural cellular self-killing. In cancerous cells, this may induce cell death at a lower level than in non-diseased cells.
  • Detection domains (2a,2b) similar to the embodiment in Figure 1 may be antibody FAb fragments against part of the Abll protein and part of the breakpoint cluster region protein (4).
  • Detection domains (2a,2b) similar to the embodiment in Figure 3C may comprise specific nucleotide sequences against part of the Abll nucleotide sequence and part of the breakpoint cluster region nucleotide sequence.
  • Activation domains (3a,3b) may comprise inactive Caspase-9 monomers.
  • the two different detection domains (2a,2b) may bind to bring two activation domains (3a,3b) together. These activation domains may interact, causing cell death. This may eliminate cancerous cells.
  • detection domains (2a,2b) may comprise antisense nucleotide strands of RNA or DNA as shown in Figures 3A-C.
  • a detection domain (2) and an activation domain (3a,3b) may be encoded for on a plasmid, as in one embodiment shown in Figure 7.
  • the plasmid may contain a sequence that enhances uptake into cells of interest.
  • the plasmid may encode detection (2a,2b) and activation (3) domains as shown in Figure 8A.
  • an activation domain may comprise a fusion of a ribonucleoprotein subunit (17a,b) and a procaspase-9 subunit.
  • a detection domain may comprise a fusion of a ribonucleoprotein-binding strand (16a,b) that is specific for the ribonucleoprotein subunit found in an activation domain and anti-sense nucleotide strand or strands that recognize nucleotide sequences produced by the cell as part of the cancer.
  • a cell containing a plasmid produces the domains, which self- assemble as shown in Figure 8B by the interaction of a ribonucleoprotein-binding nucleotide portion (16a,b) of a detection domain with a ribonucleoprotein subunit portion (17a,b) of an activation domain.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of a nucleotide strand (4) may bring the activation domains (3a,3b) into contact such that they may dimerize. The cell may apoptose when this occurs in sufficient numbers.
  • IMPs may have the distinct advantage that they may only substantially affect targeted ideotypical cancer cells. This may spare non- malignant cells from the effects of treatment. Current cancer treatments often employ drugs that disrupt cell growth, but this disruption causes severe collateral damage and side-effects. By sparing non-malignant cells, IMPs may avert the substantial side- effects of chemotherapeutics and radiation therapies. Further, chemotherapeutics and radiation therapies take a shotgun approach to proliferating cells. Newer therapies and the human immune system do attempt to create a more targeted response by recognizing such cells externally, but cancerous cells mutate rapidly and their surface antigens are often hidden.
  • IMPs may target all rather than just a subset of cancerous cells by recognizing epitopes inside the cell that are less likely to be hidden or mutable. This improved combination of specificity and efficacy of treatment may result in complete eradication of cancers in some patients.
  • IMPs may also be used to treat extracellular infections.
  • existing treatments for bacterial diseases i.e. antibiotics
  • antibiotics favor natural selection of pathogens that evolve resistance mechanisms.
  • Antibiotic resistance is a common problem, especially in environments like hospitals where antibiotics and multiple disease-causing organisms are frequently brought into contact.
  • a related problem with antibiotics is their non-specificity against virulent bacteria.
  • the term "virulent” as used herein refers to the state of any resident organism in which the organism expresses genes that help it to cause disease in a host organism. Most bacteria found in and on human beings are not virulent and are sometimes called “avirulent" strains. Unfortunately, antibiotics commonly target both virulent and avirulent organisms.
  • virulent strains including XDR Tuberculosis and MRSA— are those which have developed the strongest resistance mechanisms to existing antibiotics. Thus treating an individual with antibiotics may actually help the virulent strains to spread by removing avirulent strains that are competing for the same resources.
  • any given bacterium can switch from being avirulent to being virulent by expressing virulence genes.
  • the terms "virulence genes” and “virulence factors” as used herein refer to the genes and gene products of a resident organism that help it to cause disease in a host organism. Virulence factors may include various toxins, adhesion molecules (invasins), secretion system proteins, enzymes, capsule proteins, and immunosuppressants, to name a few. Not all strains of bacteria in a given species express or even possess virulence genes at any given time. However, all organisms that are virulent at any given time express some virulence genes. The expression of mRNA and protein from these virulence genes in high levels may be unique to virulent cells, making them ideotypical. It should be noted that these particular ideotypical cells are not human, but rather are bacterial cells.
  • detection domains (2a,2b) could comprise an antibody fragment that is specific for alpha-hemolysin or Hla (4).
  • Hla is an especially potent toxin expressed only in virulent Staph aureus and not in avirulent strains of the same bacterium.
  • An activation domain (3) could comprise a monomer of an antibacterial toxin that is effective only as a dimer.
  • the activation domains (3a,3b) may be brought into contact and dimerize. This may cause the death of virulent bacteria but spare avirulent strains and species.
  • IMPs may circumvent the resistance mechanisms of currently-resistant bacteria.
  • IMPs may help naturally select variant strains that do not express virulence factors. This means that the evolutionary pressure exerted by IMPs may exactly oppose that of classical antibiotics.
  • Classical antibiotics select for strains that can express virulence genes that directly inactivate the antibiotics, whereas IMPs may select for strains that avoid expressing virulence factors altogether.
  • IMPs may not affect avirulent commensal organisms. This means that patients using IMPs may not be susceptible to other infections as they would be when taking antibiotics. IMPs may actually give avirulent commensal organisms an evolutionary advantage over virulent organisms, keeping the patient's balance of bacterial flora in a favorable equilibrium.
  • IMPs could also be used to treat vector-bome parasites.
  • vector-born parasites refers to parasitic organisms that are transferred to humans and/or to other animals by a “vector” organism like a mosquito, a tick, or some other intermediary. These parasites can include bacteria, fungi, yeasts, and protozoa. These diseases present a particular problem in that there are often vast reservoirs of infection.
  • the term “reservoir” as used herein refers to animals other than humans in which a parasitic organism can grow and multiply and eventually be transferred by a vector to other animals or to humans. A reservoir may or may not experience a disease state from the parasite.
  • Examples of such parasites and their diseases include sleeping sickness and Chagas disease (caused by trypanosomes from tsetse flies and assassin bugs, respectively), Lyme disease (caused by Borrelia burgdorferi from ticks), and Tularemia (caused by Francisella tularensis from various arthropods), to name a few.
  • mosquitoes may be targeted.
  • a detection domain (2) may comprise an antibody fragment specific for var gene products that are expressed copiously on the surface of the protozoa.
  • An activation domain (3a,3b) may comprise an inactive subunit of an arthropod-specific toxin.
  • heterodimerization or heteromultimerization may produce an active toxin.
  • Such an embodiment may be introduced to mosquitoes by injecting it into a species that the vector mosquitoes generally feed on. As the mosquito ingests the blood of the animal, it also ingests the IMPs (1).
  • the IMPs (1) may have no epitopes (4) to bind and may therefore be inert.
  • the activation domains (3a,3b) may dimerize or multimerize to form the full toxin. In the presence of sufficient toxin, the carrier mosquito may die.
  • a similar embodiment to target malaria-carrying mosquitoes may have the same detection domain (2) but have activation domains (3a,3b) on different IMPs comprised of either a pro-toxin or a weakened enzyme to convert the toxin into its active form.
  • activation domains (3a,3b) on different IMPs comprised of either a pro-toxin or a weakened enzyme to convert the toxin into its active form.
  • the detection domains (2) of two different IMPs (la, lb) bind to adjacent epitopes (4), the pro-toxin (3a) and weak enzyme (3b) interact as may not be possible in solution because of the enzyme's relative non-specificity. Once the pro-toxin is converted in sufficient amounts, the carrier may die.
  • IMPs may target only those vectors that are actually of the carrier ideotype. Since this may be just a fraction of the vector population, it may allow the ecological niche that would be vacated by expired carriers to be filled seamlessly by non-carriers of the same species. This may minimize the environmental impact of the treatment. Further, IMPs in this example may promote the natural selection of vectors of any given species that are themselves resistant to colonization by the parasite. In some diseases, species that are resistant to colonization have already been described. These populations may be enriched in areas where carriers are selected against because of the added evolutionary advantage. The end goal of these treatments may be to eliminate transmission to human hosts. By cutting off the vector source of the parasite, the parasite may no longer be transmitted. Without such transmission, the disease may be eradicated entirely from a single region or from the world at large. Additionally, all this may be accomplished without immunization of the human population.
  • IMPs against vector- borne malaria
  • many other embodiments of IMPs targeting various parasitic diseases in various ways are possible.
  • other embodiments may be used to target malaria.
  • an embodiment may be introduced to the potential-carrier vector population by various other means including by directly feeding the vector a solution containing IMPs. These feedings may be protected such that a feeding station may not itself become colonized by the parasite.
  • the above examples should not be considered limiting.
  • autoimmune disorder refers to any disease in which the human immune system causes or exacerbates the disease. This may include classical autoimmune disorders like Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA), and Type I Diabetes as well as immunoproliferative disorders (cancers involving immune cells), transplant scenarios, allergies, and infectious diseases in which primary harm is caused by the immune system.
  • SLE Systemic Lupus Erythematosus
  • RA Rheumatoid Arthritis
  • Type I Diabetes as well as immunoproliferative disorders (cancers involving immune cells), transplant scenarios, allergies, and infectious diseases in which primary harm is caused by the immune system.
  • B and T cells of the immune system are both major and unique cells in the human body. They express a wide variety of receptors called antigen receptors, which bind various antigens throughout the body.
  • antigen receptor refers to receptors on the B cell that are also often referred to as the B cell receptor (BCR), membrane-bound Ig (mlg), and antibodies (Ab) as well as to receptors on the T cell that are also referred to as the T cell receptor (TCR).
  • BCR B cell receptor
  • mlg membrane-bound Ig
  • TCR T cell receptor
  • Each clonal population of B or T cells only expresses one antigen receptor that is specific to a particular antigen.
  • clonal refers to a group of cells that come from the same parent B or T cell. Clonal cells may be cells of a single ideotype, meaning cells that express similar epitopes (e.g. the antigen receptor).
  • FIG. 5 shows a detection domain (2) linked to an activation domain (3) by a linker (10) according to one embodiment.
  • a detection domain (2) may be an antigen for which a clonal (and ideotypical) subpopulation of cells expresses a particular antigen receptor.
  • An activation domain (3) may be FasL. FasL is a molecule that, when trimerized, may interact with the receptor Fas on the surface of nucleated cells to induce downstream signals that cause the death of the cell.
  • FIG. 6 shows the binding of a plurality of Ideotypically Modulated Pharmacoeffectors (1) to the surface antigen receptors (4) of ideotypical disease- causing B cells (11) according to one embodiment.
  • a detection domain (2) may be an antigen for which a clonal, ideotypical subpopulation of cells expresses a particular antigen receptor and an activation domain (3) may be FasL.
  • the detection domains (2) may bind the antigen receptors (4) of the disease-causing cells (11).
  • the FasL activation domains (3) trimerize.
  • FasL has trimerized, it may interact with the receptor Fas (12) on the surface of the cells to induce downstream signals. These downstream signals may cause the death of the cell.
  • the malignant B or T cell population is derived from a single cell. This means that the malignant cell population is clonal and each of these cells expresses the same or closely-related antigen receptor (i.e. has the same ideotype).
  • the embodiment's detection domain (2) may be an antigen for which the antigen receptors (4) of cancerous cells are specific.
  • an ELISA screen of potential antigens may elucidate which antigen this should be.
  • the embodiment's activation domain (3) may be FasL, which is a ligand that— when trimerized— may interact with the Fas receptor found on all nucleated cells' surfaces.
  • the targeted antigen receptors (4) may interact with IMPs on ideotypical disease-causing cells through the detection domain (2). This may bring a plurality of activation domains (3) into close proximity and they may trimerize. As a result of this trimerization, these activation domains may interact with the Fas receptor (12) on the surface of said cells to cause downstream signaling that induces apoptosis in the cell. The death of these cells may clear the disease.
  • an antibody on the surface of said B cells may actually provide the epitopes (4) to which the detection domains (2) bind.
  • an antibody may be part of the embodiment's detection domain.
  • the detection domain (2) may comprise the antigen for which said antigen receptors (4) are specific.
  • the detection domain (2) may also comprise an antibody that is specific for the membrane-bound antibody.
  • Other IMP embodiments may be used to treat patients with classical autoimmune disorders like Systemic Lupus Erythematosus (SLE).
  • autoimmune diseases generally follow a partem of mistakes in the screening of B and T cells that are reactive to self antigens.
  • self antigen refers to proteins and other metabolites produced by the human body.
  • autoreactive When B and T cells are reactive to self antigens, such cells are termed "autoreactive.” When these cells escape the screening processes of the body, they can cause damage and interrupt important processes in the body.
  • disease-related B and T cells are autoreactive to nuclear antigens.
  • the detection domain (2) of an embodiment may comprise such nuclear antigens. These could be determined by ELISA.
  • the activation domain (3) of an embodiment may comprise FasL as in some previous embodiments.
  • a plurality of IMPs (1) would bind the antigen receptors (4) of the autoreactive cell ideotype (11). This would bring the activation domains (3) of multiple IMPs (1) together to trimerize. Trimerized activation domains (3) may interact with the Fas receptor (12) on said cells, inducing apoptosis of the cell. Since the epitope (i.e. antigen receptor specific for the detection domain) may not be present on the surface of cells that do not contribute to disease, these cells may be spared. In some autoimmune disorders, multiple ideotypes may be present and may be targeted with separate embodiments.
  • T cell antigen receptors differ from B cell receptors (BCRs) in that they recognize an antigen in the context of an MHC molecule on other cells.
  • BCRs B cell receptors
  • some adjustment in the detection domain may be necessary to accommodate this difference. This may include adding an MHC motif to the detection domain (2).
  • detection domains (2a,2b) may comprise antisense nucleotide strands of RNA or DNA as shown in Figures 3A-C that are specific for sequences found in autoimmune B and/or T cells.
  • Autoimmune B and T cells have unique nucleotide sequences as a result of viral changes or somatic recombination.
  • the term "somatic recombination” as used herein refers to the V(D)J recombination that takes place during B and T cell receptor maturation. These unique sequences are an ideotypic change that could be exploited using IMPs with complementary nucleotide strands.
  • a detection domain could be simply reverse-engineered from these unique sequences.
  • a detection domain (2) and an activation domain (3a,3b) may be encoded for on a plasmid, as in one embodiment shown in Figure 7.
  • the plasmid may contain a sequence that enhances uptake into cells of interest.
  • the plasmid may encode detection (2a,2b) and activation (3) domains as shown in Figure 8A.
  • an activation domain may comprise a fusion of a ribonucleoprotein subunit (17a,b) and a procaspase-9 subunit.
  • a detection domain may comprise a fusion of a ribonucleoprotein-binding strand (16a,b) that is specific for the ribonucleoprotein subunit found in an activation domain and anti-sense nucleotide strand or strands that recognize nucleotide sequences as discussed above in the target cells.
  • a cell containing a plasmid produces the domains, which self-assemble as shown in Figure 8B by the interaction of a ribonucleoprotein-binding nucleotide portion (16a,b) of a detection domain with a ribonucleoprotein subunit portion (17a,b) of an activation domain.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of a nucleotide strand (4) may bring the activation domains (3a,3b) into contact such that they may dimerize.
  • the cell may apoptose when this occurs in sufficient numbers.
  • Other IMP embodiments may be used to combat allergies.
  • allergy and “allergies” as used herein refer to immune responses against otherwise-innocuous foreign antigens.
  • foreign antigens refers to metabolites (e.g. proteins, lipids, carbohydrates) that were not produced by the body. Innocuous foreign antigens are often found in the body and should not normally cause an immune response. B cells producing a certain class of antibody against these antigens are a major part of many of these hypersensitivities.
  • an embodiment for the purpose of clearing B cells of the allergy-causing ideotype may have a detection domain (2) that is the foreign antigen.
  • the activation domain (3) as previously, may be FasL.
  • the detection domains (2) may only bind the antigen receptors (i.e. epitopes) of ideotypical disease- causing cells (1 1).
  • the activation domains (3) may trimerize and interact with the Fas receptors (12) on the cell surface, causing apoptosis.
  • Other IMP embodiments may further be used to combat graft-versus- host disease (GVHD).
  • GVHD graft-versus- host disease
  • transplanted tissues retain their immunogenicity in the new host and cause damage to the host's existing tissues. This is generally considered the opposite of transplant rejection, wherein the transplant actually rejects the host.
  • the detection domain (2) may be the donor cells' MHC molecules.
  • the activation domain (3) may be FasL.
  • Ideotypically Modulated Pharmacoeffectors (1) may bind the antigen receptors of disease-causing (NK and NK T) cells (11).
  • the activation domains (3) may in turn trimerize and signal death through the Fas receptor (12). This may clear the donor tissue of unwanted immune cells and may avert GVHD.
  • IMP embodiments may also be used to combat transplant rejection.
  • Transplant rejection is the rejection of donor tissue by the host, which is handled by at least two ideotypical sets of host cells.
  • One subset is host T cells with antigen receptors (TCRs) that recognize the unique MHC of the donor tissue.
  • Another subset is host B cells that produce anti-donor-MHC antibodies. These antibodies are a common cause of long waits on donor waiting lists for patients receiving transplants after a previous rejection.
  • TCRs antigen receptors
  • An IMP embodiment to combat transplant rejection may have a detection domain (2) that is a mimetic of the donor's MHC molecules.
  • An activation domain (3) may be FasL.
  • the detection domain (2) may bind the antigen receptors (4), whether TCRs or BCRs, of host T and B cells.
  • the activation domains (3) may trimerize and interact with Fas receptors (12) on the offending T and B cells (11). This may signal cell apoptosis. If these subsets of host cells are ablated, a graft rejection may be avoided.
  • detection domains (2a,2b) may comprise antisense nucleotide strands of RNA or DNA as shown in Figures 3A-C that are specific for sequences found in rejecting B and/or T cells. Rejecting B and T cells have unique nucleotide sequences as a result of viral changes or somatic recombination.
  • a detection domain could be simply reverse-engineered from these unique sequences.
  • a detection domain (2) and an activation domain (3a,3b) may be encoded for on a plasmid, as in one embodiment shown in Figure 7.
  • the plasmid may contain a sequence that enhances uptake into cells of interest.
  • the plasmid may encode detection (2a,2b) and activation (3) domains as shown in Figure 8A.
  • an activation domain may comprise a fusion of a ribonucleoprotein subunit (17a,b) and a procaspase-9 subunit.
  • a detection domain may comprise a fusion of a ribonucleoprotein-binding strand (16a,b) that is specific for the ribonucleoprotein subunit found in an activation domain and anti-sense nucleotide strand or strands that recognize nucleotide sequences as discussed above in the target cells.
  • a cell containing a plasmid produces the domains, which self-assemble as shown in Figure 8B by the interaction of a ribonucleoprotein-binding nucleotide portion (16a,b) of a detection domain with a ribonucleoprotein subunit portion (17a,b) of an activation domain.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of a nucleotide strand (4) may bring the activation domains (3a,3b) into contact such that they may dimerize. The cell may apoptose when this occurs in sufficient numbers.
  • IMP embodiments may clear the malignancy and restore the patient to normal status without the side-effects of chemotherapeutic and radiation therapy methods.
  • embodiments may clear the offending autoreactive B and/or T cells at the center of the disorder, either ameliorating symptoms or curing the disease.
  • these combined embodiments may increase the likelihood of transplant success. All this may be accomplished without the immunosuppression that is a common feature of most treatments for this group of diseases. This may be especially important because many of these patients are already immunocompromised.
  • cellular diseases certain subsets of cells fail. These cells may have a given task, like providing structural strength to surrounding tissue or providing an important hormone or metabolite to other parts of the body. A number of embodiments may induce the production of metabolites in such cells to improve their function. For instance, muscular atrophy occurs in a number of diseases.
  • One embodiment may have a detection domain (2) for troponin (4), a metabolite produced in large quantities in muscle cells.
  • An activation domain (3) may be subunits of an enzyme to help assemble actin subunits, adding strength to said muscle when the activation domains are brought together.
  • Some embodiments may be effective in patients with Type II Diabetes.
  • pancreatic beta cells producing insulin are overwhelmed with the production demands incurred by high sugar and lipid levels in the blood.
  • An IMP may help by encouraging these ideotypical pancreatic beta cells or their stem cell progenitors to proliferate, or perhaps by increasing the anti-apoptotic pathways in existing beta cells. These IMPs may extend the life of these cells and may avert or postpone the onset of serious complications of disease.
  • Adipose cells may also offer a good target for some embodiments to combat lipodystrophic disease. In such diseases, adipose cells are unable to handle the volume of fats in the bloodstream. This leads to disease. By interacting with such cells specifically, their capacity may be adjusted or their survival pathways may be bolstered. Activation domains (3) of some IMP embodiments may interact with cellular pathways to decrease the efficiency of energy expenditure in the cell, depleting excessive energy stores.
  • Some current treatments for cellular diseases have non-optimal success rates as a result of collateral cell damage, especially amongst a certain subset of cells. Some embodiments may help ameliorate these side-effects in conjunction with other treatments by bolstering particular cell subsets (i.e. neurons, muscle cells, etc) from the effects of treatment.
  • cell subsets i.e. neurons, muscle cells, etc.
  • the previous examples of embodiments for various cellular applications should not be construed as limiting.
  • Each of these example embodiments and other embodiments not listed here may offer a number of advantages, mostly having to do with treatment specificity and efficacy.
  • one embodiment may have a detection domain that is specific for proteins involved in cell differentiation.
  • An activation domain (3) may be an inactive monomer of a fluorescence protein.
  • the embodiment may be introduced to a cell tissue culture.
  • the detection domain (2) may bind it.
  • Adjacent activation domains may dimerize to form an active fluorescent dimer, allowing visualization of the location and concentration of proteins in the cell as well as visualization of the cells expressing the protein.
  • a detection domain (2) may be an antisense nucleotide strand against the mRNAs (4).
  • An activation domain (3) may be inactive subunits of an enzyme that cleaves mRNAs. In this case, when the mRNAs are produced in the cell, the detection domains may bind to adjacent RNA sequences. This may bring the subunits together, which may go on to cleave mRNA and limit its expression in the cell.
  • Embodiments as may be used in emerging or weaponized diseases [0099] Various embodiments of IMPs could be used in combating emerging or weaponized diseases. For instance, various outbreaks of viral diseases have emerged as a result of human encroachment on previously-unsettled territories. Relatedly, various terrorist organizations have expressed interest and/or competence in mutating existing agents for the purpose of weaponization and attacks on urban populations.
  • Ideotypically Modulated Pharmacoeffectors offer important advantages over these treatments. For instance, the rapid mutation of emerging diseases and the artificial mutation of weaponized diseases make the production of new treatments difficult to achieve using currently-available methods. Using certain embodiments of IMPs, however, individuals combating these diseases could utilize the easily-altered specificity of detection domains in order to adapt one IMP embodiment for use against a changing threat. These embodiments could further be rapidly produced for use in emergency situations and adjusted as needed.
  • an embodiment as shown in Figure 7 may comprise a plasmid encoding various domains.
  • Detection domains (2a,2b) may comprise antisense nucleotides as shown in Figures 3A-C that are specific for the target weaponized virus.
  • the determination of these sequences for any given ideotype of weaponized virus could be accomplished using simple and well-understood techniques such as gene sequencing and subtractive hybridization. Such cells may be extracted from a patient or from cell culture for these studies. A detection domain could be simply reverse-engineered from these unique sequences.
  • a detection domain (2) and an activation domain (3 a, 3b) may be encoded for on a plasmid, as in one embodiment shown in Figure 7.
  • the plasmid may contain a sequence that enhances uptake into cells of interest.
  • the plasmid may encode detection (2a,2b) and activation (3) domains as shown in Figure 8A.
  • an activation domain may comprise a fusion of a ribonucleoprotein subunit (17a,b) and a procaspase-9 subunit.
  • a detection domain may comprise a fusion of a ribonucleoprotein-binding strand (16a,b) that is specific for the ribonucleoprotein subunit found in an activation domain and anti-sense nucleotide strand or strands that recognize nucleotide sequences as discussed above in the target cells.
  • a cell containing a plasmid produces the domains, which self-assemble as shown in Figure 8B by the interaction of a ribonucleoprotein-binding nucleotide portion (16a,b) of a detection domain with a ribonucleoprotein subunit portion (17a,b) of an activation domain.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of a nucleotide strand (4) may bring the activation domains (3a,3b) into contact such that they may dimerize.
  • the cell may apoptose when this occurs in sufficient numbers.
  • adjustments in detection domains may be easily undertaken using simple techniques to circumvent the resistance.
  • pores in, on and/or through membranes are important structural process.
  • pore and/or "pores,” as used herein, refer to any channel, tunnel, aperture, conduit, avenue, permeation, carrier, duct, canal, barrel, perforation, groove, furrow, course, passage, culvert, valve, hole, patency, enlargement, opening, orifice, outlet, cavity, or other communication between one side of any membrane and another as understood by one skilled in the art.
  • membrane and/or “membranes,” as used herein, refer to any surface, matrix, field, lining, boundary, partition, layer (whether single-layered or multiple-layered), or material separating one space from another, as understood by one skilled in the art.
  • FIG. 9 depicts an exemplary view of a membrane 19 separating a space 20a from another space 20b.
  • Membranes such as membrane 19, may restrict or reduce the movement of materials between the aforementioned spaces, such as spaces 20a and 20b, wherein the spaces, such as spaces 20a and 20b, may be separated by one or more membranes, such as membrane 19.
  • Materials that may be separated by membranes may include, but are not limited to, fluids, ions, atoms, molecules, information, solvents, solutes, potential differences, fields, or other membranes.
  • Membranes may separate one or more spaces and need not be planar (i.e.
  • Membranes may be pliable, elastic, expandable, compressible, or otherwise structurally modifiable. Further, membranes may achieve any conceivable dimension or size, including but not limited to subatomic, atomic, micromolecular, macromolecular, microscopic, and macroscopic sizes as understood by one skilled in the art.
  • Pore formation in a membrane can be accomplished by division of a membrane through the introduction of one or more subunits that operate to create an opening or other structure that causes a change in the membrane permeability for the exchange of materials between one space and another space that would otherwise be inhibited or reduced by the presence of the membrane as understood by one skilled in the art. Pore formation may also be accomplished by division of a membrane through the retraction of one or more subunits to make an opening for the exchange of materials as defined above. Such pores may allow or restrict the exchange of materials based on properties of the material in question such as charge, magnetism, size, solubility, shape, conformability, affinity, or any other criteria as understood by one skilled in the art.
  • homogeneous or heterogeneous subunits may be brought together to form a structure facilitating a pore in a given membrane.
  • These subunits may be, but are not limited in reference to, monomers, subunits, or porins.
  • the terms "monomer,” “monomers,” “porin,” “porins,” and/or related terms as used herein refer to any individual units which alone do not constitute a pore but when localized with a membrane in a correct combination— i.e. oligomerization— produce a pore structure.
  • oligomerization and multimerization refer to the colocalization of subunits or monomers wherein said colocalization of molecules changes the activity or structure of said subunits. This may constitute dimerization (i.e. oligomerization or multimerization of two subunits), trimerization (i.e. oligomerization or multimerization of three subunits), tetramerization dimerization (i.e. oligomerization or multimerization of four subunits), or any other combination of subunits.
  • the number of subunits may vary in any given complex of subunits and said structures may be formed by identical or non-identical subunits of various kinds.
  • oligomerize and “multimerize” may refer to the activity of apposition of multiple subunits as understood by one skilled in the art. These activities may comprise structural formation, direct enzymatic activity, or other multimerization activity. Oligomerization may occur naturally or be induced. An artificial Ideotypically Modulated Pharmacoeffector may be induced, or otherwise activated, to oligomerize to form a structure in the presence of detectable epitopes according to one embodiment. Such a structure may be a pore.
  • Examples of naturally -produced pores include Perfringolysin O produced by Clostridium bacteria, Cytolysin A produced by E. coli bacteria, Pneumolysin produced by Streptococcus bacteria, heterogeneous Leukocidins produced by Staphylococcus aureus, the complement membrane attack complex produced by human cells, and Perforin produced by human cells.
  • These structures can be modified and/or mimicked by artificial subunits and/or artificially-expressed and/or modified protein domains to produce additional porin types.
  • an identical activation domain may be advantageous.
  • alternating or heterologous activation domains may be advantageous.
  • Examples of naturally-occurring microscopic membranes include but are not limited to cell walls, lipid bilayers, lipid layers, protein walls, lipid walls, phospholipid walls, liposomes, molecular matrices, atomic sheaths, external eukaryotic cell membranes, bacterial cell walls, bacterial cell membranes, artificial lipid bilayers, mitochondrial bilayers, endoplasmic reticulum membranes, nuclear membranes, and other membranes as understood by one skilled in the art.
  • artificial membranes include but are not limited to microsomes, fullerenes, buckminsterfullerenes, polycyclic aromatic hydrocarbons, silicon sheets, graphene, cholesterol-containing membranes, polymers, ceramics, amorphous solids, and/or other combinations thereof and other like membranes.
  • Figure 10A shows one example of an unbound Ideotypically Modulated Pharmacoeffector (1) that may be introduced into a population of cells or localized with a membrane according to one embodiment.
  • the detection domains (2a, 2b) of one or more embodiments of IMPs (1) have affinity for adjacent or otherwise-apposed epitopes on a predetermined target antigen, such as target antigen (4) shown in Figure 10B, when present in the cell or near a membrane of a particular ideotype.
  • target antigen (4) shown in Figure 10B, when present in the cell or near a membrane of a particular ideotype.
  • epitopes as further used herein refers to any part of a molecule of interest with which a detection domain, as defined above, interacts or binds.
  • This may include, for example, single-stranded nucleotide sequences, double-stranded nucleotide sequences, lipids, proteins, carbohydrates, combinations of materials, and such epitopes as may be detected by naturally-occurring or artificially -produced antibodies, antibody fragments, lectins, complement proteins, or nucleotide sequences according to one embodiment.
  • the terms "ideotype,” “ideotypical,” and “ideotypically” as further used herein refer to the uniqueness or differential expression of the set of antigens (and their concomitant epitopes) expressed in or on a subset of cells or membranes.
  • antigen as further used herein refers to any molecule of interest with epitopes.
  • the detection domains (2a,2b) may be identical or different, depending on the application. For instance, in detecting antigens (4) with nucleotide sequences, detection domains (2a,2b) of antisense nucleotide sequences that are complementary to adjacent sequences may be advantageous.
  • the binding of more than one Ideotypically Modulated Pharmacoeffector (1) to adjacent epitopes results in the oligomerization or multimerization of the activation domains (3) as shown in Figure IOC.
  • the terms "oligomerization” and multimerization” as further used herein refer to the colocalization of molecules, whether homologous or heterologous, changing the activity of said molecules. This may constitute dimerization (i.e.
  • oligomerization or multimerization of two subunits trimerization (i.e. oligomerization or multimerization of three subunits), tetramerization dimerization (i.e. oligomerization or multimerization of four subunits), or any other combination of subunits.
  • the number of subunits may vary in any given activated complex of Ideotypically Modulated Pharmacoeffectors. These can be referred to broadly as “multimers” or “oligomers” and are known to one skilled in the art.
  • the terms “oligomerize” and “multimerize” further refer to the activity of apposition of multiple subunits and is understood by one skilled in the art. These activities may comprise the formation of a structure, direct enzymatic activity, or other multimerization activity.
  • activation domains may be identical or different, depending on the application.
  • an activation domain (3) for inducing cell death may include an inactive modified porin monomer, which— when multimerized— converts itself to an active form in a cell membrane to produce a structure, such as a pore as defined above and/or understood by one skilled in the art.
  • a structure such as a pore as defined above and/or understood by one skilled in the art.
  • Figure 10D shows one example of binding of multiple adjacent ideotypical epitopes to the detection domains of multiple Ideotypically Modulated Pharmacoeffectors to form a structure according to one embodiment.
  • an identical activation domain may be advantageous.
  • alternating or heterologous activation domains may be advantageous.
  • modified porin monomers, domains, motifs, or other subunit types or artificial porin structures may have a number of advantages in Ideotypically Modulated Pharmacoeffectors. Some of these advantages are discussed here, but other advantages may exist.
  • One advantage may include that porin-based Ideotypically Modulated Pharmacoeffectors may facilitate improved cell uptake versus other Ideotypically Modulated Pharmacoeffectors according to one embodiment.
  • Another advantage may include non-enzymatic activation of the activation domains according to one embodiment.
  • Another advantage may include greater specificity of activation according to one embodiment.
  • Figure 11 A shows one example of an Ideotypically Modulated Pharmacoeffector (1) that may be introduced into a population of cells and/or localized near a cell membrane according to one embodiment.
  • many detection domains (2a,2b) may be present in any given Ideotypically Modulated Pharmacoeffector (1).
  • many activation domains (3) may be present in any given Ideotypically Modulated Pharmacoeffector (1).
  • detection domains (2a,2b) may comprise antisense nucleotide strands.
  • activation domains (3) may perform substantially the same functions as the activation domain shown in Figures 1 A- 3C and/or 10A-10D. These elements may be conjugated (2a,3 and 2b,3) to form IMPs (la, lb, etc) by carbodiimides or by some other functional group or linker in a chemical process.
  • epitopes (4) may comprise nucleotide strands that may be unique to target cells of interest.
  • these strands may comprise a nucleotide sequence introduced by a virus as it infects a cell.
  • the detection domains (2a,2b) may be antisense nucleotide sequences that are complementary to that introduced by the virus.
  • the epitope (4) may be expressed in a particular target cell type (i.e. ideotype), whether viral, cancerous, or otherwise of interest. In certain other embodiments, the epitope may be expressed in varying levels in a particular target cell type or ideotype.
  • One or more detection domains of various kinds may be attached to a single activation domain, as depicted here. Similarly, one or more activation domains may be attached to a single detection domain. In addition, a plurality of detection domains may be attached to a plurality of activation domains. Further, a single entity including one or more activation domains or one or more detection domains may be produced without requiring separate linkage. For instance, a protein containing both activation domain or domains and detection domain or domains may be produced as known to one skilled in the art.
  • FIG 12A shows one example of Ideotypically Modulated Pharmacoeffectors (1) that have been introduced to a membrane (5) of a cell (18).
  • a membrane may be associated with a cell in some embodiments.
  • Ideotypically Modulated Pharmacoeffectors may span the cell surface membrane. In other embodiments, they may be found inside the cell.
  • the Ideotypically Modulated Pharmacoeffectors in one embodiment do not multimerize optimally as understood by one skilled in the art.
  • Figure 12B shows one example of Ideotypically Modulated Pharmacoeffectors (1) that have been introduced to a naturally-occurring cell (18) which has one or more naturally-occurring epitopes (4) present within the cell.
  • these epitopes may comprise nucleotide sequences introduced or produced by a virus in the cell.
  • these epitopes may comprise proteins produced naturally in a cancerous cell.
  • these epitopes may comprise lipids produced naturally by a subset of cells.
  • these epitopes may be on or near the outer surface of a cell.
  • these epitopes may comprise cell surface receptors produced naturally by a subset of cells.
  • the embodied cell (18) may be archetypal of a population of similar cells.
  • the Ideotypically Modulated Pharmacoeffectors (1) in one embodiment multimerize readily to form a structure in a manner depicted in Figure 12C.
  • the structure may comprise a pore.
  • Detection domains (2a,2b) may comprise antisense nucleotide strands of RNA or DNA as shown in Figures 10A-D.
  • a detection domain (2a,2b) of an embodiment may comprise antisense nucleotide strands that are complementary to adjacent conserved sequences of ideotypical viral nucleotides (4). These strands could be manufactured by using PCR or by transfecting bacteria to produce them.
  • the activation domain (3) of an embodiment may be a modified inactive monomer of a porin such as Perfringolysin O.
  • a detection domain (2) and an activation domain (3) may be conjugated together through a benzaldehyde group interacting to an N-terminal histamine residue on the activation domain using a modified Pictet-Spengler reaction in glacial acetic acid.
  • the binding of the detecting anti-sense strands (2a,2b) to adjacent portions of an antigenic determinant HIV-specific nucleotide strand (4) may bring the activation domains (3) into contact such that they may multimerize.
  • the cell may apoptose when this occurs at the cell membrane and/or at another lipid bilayer (e.g. mitochondria, endoplasmic reticulum, phagolysosome, etc). Apoptosis may more readily occur when multimerization occurs in the cell in significant numbers.
  • lipid bilayer e.g. mitochondria, endoplasmic reticulum, phagolysosome, etc.
  • the Ideotypically Modulated Pharmacoeffectors multimerize in the presence of specific epitopes on specific antigens (4) to form a structure.
  • the activation domains of the Ideotypically Modulated Pharmacoeffectors comprise modified porin monomers. These porin monomers may readily form a pore structure on oligomerization, as is clear to one skilled in the art. In one embodiment, this oligomerization may cause pore formation and may facilitate the destruction of the cell, particularly when pore formation occurs in significant numbers. In still further embodiments, targeted porin monomers may further be modified to more readily obviate natural oligomerization as understood by one skilled in the art.
  • pore formation may produce a stereospecific channel to allow the transportation of specific ions through a membrane.
  • a channel may have substantially the same effect of facilitating apoptosis of a cell.
  • the stereospecificity of the channel may permit selective communication through the membrane of only one (or a subset) of stereoisomerically inclined molecules and/or molecular structures.
  • pore formation may allow the selective uptake of pharmaceuticals designed to target specific cells on the basis of ideotype.
  • Diphtheria toxin B monomers may be modified to allow selective uptake of an attached growth factor in adipocytes.
  • Diphtheria toxin B monomer-based IMPs may be targeted to activate in the presence of protein Myf5 that is found in brown adipose tissue and not white adipose tissue. This activation may provide a pore for the selective uptake of a growth factor in order to promote brown adipose tissue development.
  • Such an embodiment may provide the advantage of increasing energy expenditure and inducing weight loss in a body.
  • pore formation may provide for the selective uptake of chemotherapies to fight cancer.
  • a modified Diphtheria toxin B monomer may be modified to allow selective uptake of an attached cytotoxic chemotherapeutic moiety.
  • Such an embodiment may provide the advantage of minimizing off-target collateral damage and side-effects associated with systemic chemotherapy use.
  • detection domains (2a,2b) could comprise anti-sense nucleotide sequences specific for the gene for alpha-hemolysin or Hla (4).
  • the Hla gene is expressed only in virulent Staph aureus and not in avirulent strains of the same bacterium.
  • An activation domain (3) could comprise a monomer of a porin that is effective only as a multimer. If adjacent epitopes on alpha-hemolysin are detected by the detection domains (2a,2b) inside Staph cells of the virulent ideotype, the activation domains (3a,3b) may be brought into contact and dimerize.
  • a detection domain (2) that is a lectin.
  • Lectins are found in nature and are capable of binding specific sugars, as are frequently expressed on the membranes of specific bacteria. Some lectins, for instance, are quite specific to the sugars expressed on species of Chlamydia.
  • the activation domain (3) in this embodiment may include various antimicrobials.
  • effector molecules such as effector molecules (9) depicted in Figure 4, resident in the pouch or cell (18) may include antimicrobial porins.
  • porins On binding of lectins (2) to the specific sugars found on an invading bacterial cell wall in ideotypical cells, porins may be formed to digest bacterial cell barriers.
  • detection domains (2) that are anti-sense artificial nucleotide sequences specific for a mutated form of the p53 gene (4) found in many cancers.
  • An activation domain (3) may comprise inactive modified C9 protein.
  • the detection domains (2) of multiple IMPs (1) may bind mutant cancer-associated p53 mRNA (4).
  • Their activation domains (3) may oligomerize to cause the formation of a Membrane Attack Complex (i.e. a form of porin that can disrupt phospholipid bilayers of target cells, as known to one skilled in the art). When this activity reaches significant levels, the cell may apoptose.
  • a detection domain (2) may comprise an antibody fragment specific for PSA (prostate specific antigen). While PSA is ideotypical of all prostate cells rather than just cancerous cells, it may be more effective in coping with prostate cancer after it has been diagnosed.
  • An activation domain (3) may comprise inactive modified Pneumolysin. If administered in smaller amounts, the embodiment may lower the threshold of natural cellular self-killing. In cancerous cells, this may induce cell death at a lower threshold level than in non- diseased cells.
  • detection domains (2a,2b) may comprise antisense nucleotide strands of RNA, DNA, or some artificial nucleotide strands as shown in Figures 11A-D.
  • a detection domain (2) and an activation domain (3 a, 3b) may be encoded for on a plasmid, as in one embodiment shown in Figure 7.
  • the plasmid may contain a sequence that enhances uptake into cells of interest.
  • the plasmid may encode detection (2a,2b) and activation (3) domains.
  • an activation domain may comprise a fusion of a ribonucleoprotein subunit (17a,b) and a leukocydin subunit.
  • a detection domain may comprise a fusion of a ribonucleoprotein-binding strand (16a,b) that is specific for the ribonucleoprotein subunit found in an activation domain and anti-sense nucleotide strand or strands that recognize nucleotide sequences produced by the cell as part of the cancer.
  • a cell containing a plasmid produces the domains, which self-assemble as shown in Figure 8B by the interaction of a ribonucleoprotein- binding nucleotide portion (16a,b) of a detection domain with a ribonucleoprotein subunit portion (17a,b) of an activation domain.
  • the binding of the anti-sense strands (2a,2b) to adjacent portions of a antigenic determinant nucleotide strand (4) may bring the activation domains (3a,3b) into proximity or contact such that they may dimerize.
  • the cell may apoptose, particularly when this dimerization occurs in significant numbers.
  • a particular embodiment to target malaria-carrying mosquitoes may have any detection domains (2) but have activation domains (3a,3b) comprising a modified alpha-Hemolysin porin subunit.
  • the detection domains (2) of multiple IMPs (1) bind to adjacent epitopes (4), the alpha-Hemolysin subunit activation domains are brought together to oligomerize and may cause damage to the host— in this example, the malaria-carrying mosquitos. This may lead to a severe selective disadvantage to the carrier state, causing mosquitoes to evolve toward the non-carrier state and preventing disease.
  • Ideotypically Modulated Pharmacoeffectors (1) may be programmed against a specific B-cell receptor gene found exclusively or more frequently in autoimmune-associated B cells.
  • Activation domains may comprise modified porin subunit Cytolysin A.
  • oligomerization is not favored due to the absence of the specific gene.
  • autoimmune B cells in a given situation may express such a gene to produce mRNA as understood by one skilled in the art.
  • multiple IMPs are induced to oligomerize due to detection domain binding. In the embodiment shown in Figure 12D, this causes the formation of pores and facilitates the destruction of the specific autoimmune B cells.
  • Perfringolysin O monomers (21 7) can be modified to remove their natural, spontaneous oligomerizing activity.
  • Figure 13A shows one example of part of a model porin monomer of Perfringolysin O according to one embodiment.
  • Perfringolysin O is produced as a monomer by the bacterium Clostridium perfringens. Pictured are the beta sheets (22) and hairpin loops (23) of Domain 3 of Perfringolysin O in its soluble, monomelic form that occurs in nature. The remainder of Perfringolysin O monomer is depicted as an ellipsis, three consecutive dots.
  • Naturally-occurring Perfringolysin O has the following protein sequence (NCBI accession AAA23271) as understood by one skilled in the art:
  • the normal steps for Perfringolysin O (PFO)-mediated cell cytotoxicity are a) membrane association of the monomer, b) hairpin insertion, and c) oligomerization to cause cytolysis.
  • Membrane association of the monomer may be initiated in PFO by a tryptophan-rich loop E458-R468 on Domain 4, which is a common feature of many family members, including Streptolysin O. This motif does not cross the entire lipid membrane.
  • Hairpin insertion may involve the residues depicted in Figure 13 A. Hairpin 1 and Hairpin 2 normally form alpha helices at the base of PFO Domain 3 (23).
  • Hairpin 1 may comprise residues Q191-A213, whereas Hairpin 2 may comprise residues K295-N305.
  • these alpha helices extend into a lipid bilayer to extend residues into the cytosol as shown in exemplary nature in Figure 13B (25).
  • the residues that may be extended into the cytosol is E204. This spanning may occur even if the PFO monomer is not associated with other monomers. In their natural state, when localized with a membrane, PFO monomers may oligomerize by interaction of residues found in various parts of the monomer (22).
  • FIG. 13B shows the natural oligomerization of a multiplicity of porin monomers (21a, 21b, ...) in the lipid bilayer (19 ...) of a given cell (18 ). Also depicted are individual amino acids known to be involved in oligomerization of Perfringolysin O monomers in nature (24) and individual amino acids that may be functionalized for use in Ideotypically Modulated Pharmacoeffectors (25) according to one embodiment. As previously, parts of the embodiments not shown directly in the figure are indicated by ellipses comprising three consecutive dots.
  • residue E204 the leading cytosolic tip of Hairpin 1 (25)— may be modified to a cysteine (E204C) and be labeled with a functional group without severely impacting protein activity.
  • This functional group may comprise one or more detection domains (2) as shown in Figure 13C according to one embodiment.
  • Figure 13C shows an Ideotypically Modulated Pharmacoeffector (1) that includes an activation domain (3, 3) comprising part or all of a porin with modification of amino acids involved in oligomerization (24) and/or functionalization for the attachment of one or more detection domains (2) according to one embodiment.
  • the IMP monomer (1) is embedded in the cell membrane (19) of the cell (18, ...) according to one embodiment.
  • oligomerization may be accomplished through a maleimide linker, or other reactive cross-linker, as known to one skilled in the art.
  • detection domains (2) may be added anywhere along an activation domain (3) as depicted in Figure 11 A.
  • residues involved in oligomerization (24) may be modified as shown in Figure 13C to prevent, or otherwise significantly hinder, spontaneous oligomerization.
  • residue Y181 may be modified to an alanine to prevent, or otherwise significantly hinder, spontaneous oligomerization (Y181A).
  • residue N331 may be modified to a glutamate to prevent, or otherwise significantly hinder, spontaneous oligomerization (N331E).
  • both of these residues may be modified to prevent, or otherwise significantly hinder, spontaneous oligomerization.
  • Figure 13D shows Ideotypically Modulated Pharmacoeffectors comprising some or all of the modifications in Figure 13C that have oligomerized or oligomerized more readily as a result of the binding of detection domains (2) to antigens with epitopes found in the cell (4) according to one embodiment.
  • This oligomerization may form a structure according to one embodiment. Again, as previously, parts of the embodiments not depicted directly in the figure are indicated by ellipses comprising three consecutive dots. These modifications may be done in combination with other modifications, some of which modifications will also prevent, or otherwise significantly hinder, spontaneous oligomerization. In some embodiments, other residues not involved in functionalization or preventing/hindering oligomerization may be modified to confer additional advantages.
  • cysteines from the remainder of the sequence may be removed in order to allow specificity of functionalization at site E204C.
  • Swaths of sequence that may not be necessary to pore formation or attachment may also be removed to confer additional advantages.
  • Related steps are known to those skilled in the art and none of the above embodiments, methods, combinations, modifications, or explanations should be considered limiting. These may also be mixed and matched or otherwise combined for varying advantages as is clear to one skilled in the art.
  • a pouch-based embodiment in Figure 4 has been mentioned in an example of an embodiment that may help treat a Chlamydia infection
  • a pouch-based embodiment may also be employed against ideotypical carrier vectors of a disease like malaria.
  • an embodiment with a nucleotide sequence for a detection domain (2) has been mentioned in an example of an embodiment that may help treat an HIV infection
  • an embodiment with a nucleotide sequence detection domain (2) may be used in treating a cancer with characteristic or enriched nucleotide sequences (4) in the cell.
  • the introduction of IMPs into a population of cells may be intra-body.
  • instructions for manufacturing IMPs may initially be introduced into a body. Then, when such IMPs are produced by cells within the body, the IMPs may be introduced to a population of cells within the body.
  • the IMPs may be produced external of a body and introduced into the body.
  • the IMPs may be introduced into a population of cultured cells. Further, IMPs may be introduced to a membrane of any kind.

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Abstract

Dans un mode de réalisation, un procédé comprend l'introduction d'une pluralité d'effecteurs pharmaceutiques à modulation idéotypique (IMP) dans une population de cellules. Chaque IMP peut comprendre un domaine de détection et un domaine d'activation. Un ou plusieurs épitopes sont liés par ledit domaine de détection. Le domaine d'activation est activé en réponse à la liaison. Ces applications peuvent comprendre, entre autres, des infections virales, d'autres infections intracellulaires, des cancers, des maladies à transmission vectorielle, des maladies auto-immunes, des maladies cellulaires, le renforcement cellulaire et la recherche.
PCT/US2016/014751 2015-01-23 2016-01-25 Effecteurs pharmaceutiques à modulation idéotypique pour traitement cellulaire sélectif WO2016118956A1 (fr)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014004694A1 (fr) * 2012-06-26 2014-01-03 Massachusetts Institute Of Technology Dissimulation réversible de protéines formant des pores pour distribution macromoléculaire
US20140057351A1 (en) * 2010-05-31 2014-02-27 Imperium Biotechnologies, Inc. Ideotypically modulated pharmacoeffectors for selective cell treatment

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140057351A1 (en) * 2010-05-31 2014-02-27 Imperium Biotechnologies, Inc. Ideotypically modulated pharmacoeffectors for selective cell treatment
WO2014004694A1 (fr) * 2012-06-26 2014-01-03 Massachusetts Institute Of Technology Dissimulation réversible de protéines formant des pores pour distribution macromoléculaire

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
RAMACHANDRAN ET AL.: "Membrane-dependent conformational changes initiate cholesterol- dependent cytolysin oligomerization and intersubunit bold beta-strand alignment.", NATURE STRUCTURAL & MOLECULAR BIOLOGY, vol. 11, no. 8, August 2004 (2004-08-01), pages 697 - 705 *

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