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Penam

Penam is a bicyclic β-lactam compound characterized by a four-membered β-lactam ring fused to a saturated five-membered thiazolidine ring, with the systematic name 4-thia-1-azabicyclo[3.2.0]heptan-7-one, serving as the fundamental core structure of the penicillin class of antibiotics.^[1]^[2] This rigid bicyclic system, typically featuring a 5R configuration at the fusion site unless otherwise specified, imparts significant ring strain that contributes to the reactivity essential for its antibacterial mechanism.^[1]^[3] Penams function by mimicking the D-alanyl-D-alanine terminus of bacterial peptidoglycan precursors, thereby irreversibly inhibiting penicillin-binding proteins (PBPs) and disrupting cell wall cross-linking, which leads to bactericidal activity against Gram-positive and certain Gram-negative bacteria.^[4] The penam nucleus, with the molecular formula C₅H₇NOS, forms the basis for numerous derivatives, including natural penicillins like penicillin G and synthetic analogs such as ampicillin, where modifications at the 6-amino position of the β-lactam ring modulate spectrum, stability, and resistance profiles.^[5]^[4] In addition to direct antibiotics, penam-based sulfones like sulbactam and tazobactam act as β-lactamase inhibitors, protecting partner β-lactam drugs from enzymatic degradation and extending their utility against resistant pathogens.^[2] These compounds are produced semisynthetically from 6-aminopenicillanic acid (6-APA), a key fermentation-derived intermediate, highlighting penam's central role in the development of β-lactam antibiotics since the mid-20th century.^[4]

Definition and Structure

Core Bicyclic System

The penam core constitutes the fundamental bicyclic scaffold of the penicillin class of β-lactam antibiotics, characterized by the fusion of a four-membered β-lactam ring to a five-membered thiazolidine ring. This architecture forms the systematic name 4-thia-1-azabicyclo[3.2.0]heptan-7-one, where the [3.2.0] designation in the von Baeyer nomenclature indicates the bridge lengths connecting the bridgehead atoms in the bicyclic framework.^[1] The thiazolidine ring incorporates a sulfur atom and a nitrogen atom as heteroatoms, while the β-lactam ring features the characteristic amide functionality essential to the system's reactivity.^[6] The molecular formula of the unsubstituted penam core is C₅H₇NOS, corresponding to a molar mass of 129.18 g/mol.^[7] In the conventional numbering system for penams—which differs from the strict von Baeyer system but is widely adopted in chemical literature—the sulfur atom occupies position 1 within the thiazolidine ring, the bridgehead nitrogen is at position 4, and the carbonyl carbon of the β-lactam is at position 7.^[1] The ring fusion occurs between positions 4 (nitrogen) and 5 (carbon) of the thiazolidine with the β-lactam, creating a strained, cis-fused bicyclic array. A textual representation of the core topology highlights the connectivity: the thiazolidine spans S¹–C²–C³–N⁴–C⁵, while the β-lactam closes via N⁴–C⁷(=O)–C⁶–C⁵, with the geminal dimethyl substitution at C² typical of natural derivatives.^[8] This core structure provides the foundational template for penicillin derivatives, where substituents at positions 2, 3, and 6 modulate biological activity while preserving the essential bicyclic integrity.^[1]

Nomenclature and Stereochemistry

The penam core structure is systematically named as (5R)-4-thia-1-azabicyclo[3.2.0]heptan-7-one according to IUPAC nomenclature, where the bicyclic system is numbered with the nitrogen at position 1, sulfur at 4, and the β-lactam carbonyl at 7; variations in literature may use alternative numbering, but the 4-thia-1-aza designation is standard.^[1]^[5] In natural penams, such as those found in penicillins, the stereochemical configuration is predominantly (2S,5R,6R), with the 5R designation specifying the absolute configuration at the fusion site between the β-lactam and thiazolidine rings; this 5R fusion is essential for the compound's biological activity as an antibiotic.^[9]/05:_Stereochemistry_at_Tetrahedral_Centers/5.13:_Chemistry_MattersChiral_Drugs) The β-lactam ring is cis-fused to the thiazolidine ring at the 5,6-bond, resulting in a rigid bicyclic framework that constrains the overall geometry.^[2] The thiazolidine ring adopts an envelope conformation, with one atom (typically C3 or S1) displaced out of the plane formed by the other four, enabling limited pseudorotation between C2- and C3-puckered states while maintaining structural integrity.^[10]^[11] Stereoisomers of the penam core, including those with trans fusion at the 5,6-position or inverted configurations at C2 or C6 (e.g., 2R,5S,6S), are rare in natural sources due to the high stereospecificity of the biosynthetic pathway, which selectively forms the (2S,5R,6R) isomer during β-lactam ring closure.^[12]^[13] This stereospecificity ensures the bioactive conformation required for enzyme inhibition.

History and Discovery

Isolation from Penicillin

The discovery of penicillin occurred in 1928 when Scottish bacteriologist Alexander Fleming observed antibacterial activity from a mold contaminant, identified as Penicillium notatum, in his laboratory cultures of Staphylococcus bacteria at St. Mary's Hospital in London.^[14] Fleming noted that the mold secreted a substance that inhibited bacterial growth around it, terming it "penicillin" after further testing, though initial efforts to purify and characterize it were limited.^[15] During the early 1940s, amid World War II demands for new treatments against bacterial infections, Australian pathologist Howard Florey, German-born biochemist Ernst Chain, and biochemist Norman Heatley at the University of Oxford undertook systematic efforts to purify and characterize penicillin from Penicillium fermentations.^[14] Their work, starting in 1939, involved developing extraction methods using solvents and acids to isolate crude penicillin, enabling the first animal trials in 1940 that demonstrated its efficacy against streptococcal infections in mice.^[16] These purification advances scaled production for limited human trials by 1941, treating wounded soldiers, and contributed to the recognition of penicillin's core β-lactam functionality through ongoing degradation and analytical studies.^[14] In 1957, researchers at Beecham Research Laboratories in England isolated 6-aminopenicillanic acid (6-APA), the fundamental penam nucleus of penicillin, by enzymatic hydrolysis of penicillin G using penicillin acylase from microbial sources during fermentation.^[17] This breakthrough revealed the bicyclic penam core, consisting of a β-lactam ring fused to a five-membered thiazolidine ring, through targeted degradation of the acyl side chain. The isolation of 6-APA enabled the synthesis of semi-synthetic penicillins by attaching varied side chains, addressing limitations like penicillinase resistance in natural forms.^[17] That same year, 1957, marked the commercialization of 6-APA production via optimized submerged fermentation of high-yielding Penicillium chrysogenum strains, followed by enzymatic cleavage, allowing industrial-scale output for derivative development.^[18]

Structural Elucidation

The structural elucidation of penam, the core bicyclic β-lactam-thiazolidine system of penicillin, began in the early 1940s amid wartime secrecy and limited material availability. Initial chemical degradation studies, led by Edward P. Abraham and Ernst B. Chain at Oxford University starting in 1942, involved acid and alkaline hydrolysis of penicillin salts, yielding key fragments such as penicillamine (a thiazolidine derivative) and penilloaldehyde. These degradations revealed a sulfur-containing nucleus with an amide linkage, suggesting a fused ring system, though the exact configuration remained elusive due to the molecule's instability and the scarcity of pure samples (often less than 500 mg at high purity).^[19] Early structural proposals faced significant challenges, including misattributions to alternative frameworks like the thiazolidine-oxazolone structure advocated by Robert Robinson and John Cornforth in 1944, which accounted for some degradation products but failed to explain penicillin's non-basic nature and reactivity. Collaborative efforts across Oxford, Merck, and other institutions, formalized in 1944 under Anglo-American agreements, resolved these ambiguities by October 1945 through integrated chemical analysis; Abraham's 1943 proposal of a β-lactam ring fused to a thiazolidine was accepted as it aligned with hydrolysis patterns and excluded oxazolone inconsistencies, such as mismatched pK values for model compounds.^[19]^[20] Definitive confirmation came from X-ray crystallography in the late 1940s, pioneered by Dorothy Crowfoot Hodgkin and Barbara Rogers-Low using sodium, potassium, and rubidium salts of benzylpenicillin. Despite challenges like non-isomorphous crystals and limited computational resources, Patterson and Fourier methods mapped the electron density, pinpointing the sulfur atom and verifying the strained β-lactam-thiazolidine fusion with precise bond distances; this work, completed by 1945 but published in 1949 due to security restrictions, provided the first three-dimensional visualization of the penam core.^[21]^[20] In the 1950s, advancing spectroscopic techniques further refined bond confirmations, with early nuclear magnetic resonance (NMR) spectroscopy identifying proton environments in the thiazolidine ring and infrared (IR) spectra corroborating the β-lactam carbonyl stretch around 1780 cm⁻¹. Mass spectrometry emerged to analyze fragmentation patterns, supporting the molecular formula C₁₆H₁₉N₂O₄S for benzylpenicillin derivatives. The total synthesis of penicillin V by John C. Sheehan and Kenneth R. Henery-Logan in 1957 at MIT provided ultimate validation, as the synthetic product matched natural penam's optical rotation, chromatographic behavior, and antibacterial activity, solidifying the structure without reliance on degradation alone.^[22]^[23]

Synthesis

Biosynthetic Pathways

The biosynthesis of the penam core, the bicyclic β-lactam structure central to penicillins, occurs primarily in filamentous fungi such as Penicillium chrysogenum through a multi-enzymatic pathway that assembles and cyclizes specific amino acids.^[24] The pathway initiates with the condensation of three amino acids: L-α-aminoadipate (derived from lysine catabolism), L-cysteine, and L-valine (which undergoes epimerization to D-valine during the process).^[25] This tripeptide intermediate, δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine (ACV), is formed by the multifunctional enzyme ACV synthetase, a non-ribosomal peptide synthetase that activates and links the amino acids without ribosomal involvement.^[26] Subsequently, isopenicillin N synthase (IPNS), an iron-dependent oxygenase, catalyzes the oxidative cyclization of ACV to produce isopenicillin N, the first compound containing the penam core, through a four-electron oxidation process involving molecular oxygen and a reductant like glutathione.^[27] Finally, isopenicillin N acyltransferase exchanges the L-α-aminoadipoyl side chain of isopenicillin N with phenylacetic acid (or other acyl groups) to yield active penicillins such as penicillin G.^[28] The key reaction forming the penam ring can be represented as:

\delta\text{-(L-}\alpha\text{-aminoadipyl)-L-cysteinyl-D-valine (ACV)} \xrightarrow{\text{IPNS}} \text{isopenicillin N}

This cyclization establishes the characteristic 6-aminopenicillanic acid (6-APA) nucleus via β-lactam ring closure between the cysteinyl carboxyl and the valinyl amino group, with the thiazolidine ring forming concurrently.^[29] In Penicillium species, the biosynthetic genes are organized in a compact cluster on a single chromosome, facilitating coordinated regulation. The ACV synthetase is encoded by pcbAB, a large gene producing a multidomain protein, while pcbC codes for IPNS; these are followed downstream by penDE, which encodes the acyltransferase as a bifunctional protein handling both epimerization and acylation.^[30] This gene cluster, spanning approximately 18 kb, is highly conserved among penicillin-producing fungi and responds to environmental cues like carbon source limitation, with glucose repression inhibiting transcription of pcbAB and pcbC.^[31] The clustered organization likely originated from horizontal gene transfer from bacterial ancestors, as evidenced by sequence similarities.^[32] Variations in the pathway occur in bacterial and other fungal producers, such as Acremonium chrysogenum (formerly Cephalosporium acremonium), where the early steps mirror those in Penicillium—ACV formation by pcbAB and cyclization to isopenicillin N by pcbC—but diverge afterward.^[33] Instead of side-chain exchange, isopenicillin N is epimerized to penicillin N, followed by ring expansion via expandase/hydroxylase enzymes (cefEF) to form the seven-membered dihydrothiazine ring of the cepham core, ultimately yielding cephalosporin C.^[34] This modification adapts the β-lactam scaffold for broader antibiotic applications while retaining the initial penam intermediate.^[35] Industrial production of penicillins leverages optimized strains of P. chrysogenum engineered for amplified gene cluster expression, enhancing yields through this natural pathway.^[28]

Chemical Synthesis Methods

The first total synthesis of a penicillin derivative, specifically penicillin V, was achieved by John C. Sheehan and Kenneth R. Henery-Logan in 1957, marking a landmark in organic synthesis for constructing the penam core. This route began with the formation of a thiazolidine ring from L-cysteine and phenoxyacetaldehyde, followed by protection and elaboration to an azetidinone intermediate. The β-lactam ring was then closed via a cyclization step involving activation of the carboxylic acid, ultimately yielding the bicyclic penam system after deprotection and side-chain installation.^[36] Semi-synthetic approaches dominate industrial production of penam-based antibiotics, starting from 6-aminopenicillanic acid (6-APA), which is obtained by enzymatic hydrolysis of natural penicillins like penicillin G using penicillin acylase. The 6-APA nucleus retains the intact penam skeleton, allowing acylation at the 6-amino position with various carboxylic acid derivatives—typically activated as acid chlorides, anhydrides, or via enzymatic coupling—to introduce diverse side chains. This method enables efficient production of analogs such as ampicillin and amoxicillin, with yields often exceeding 90% in optimized processes.^[37]^[38] Key reactions in penam synthesis include the [2+2] cycloaddition, known as the Staudinger reaction, which forms the β-lactam ring by reacting a ketene (generated from an acid chloride) with an imine derived from the thiazolidine precursor. This stereoselective cycloaddition establishes the fused ring system with control over the cis configuration at the β-lactam fusion. Thiazolidine assembly typically proceeds via imine condensation between a cysteine derivative and an aldehyde, followed by intramolecular nucleophilic attack to close the five-membered ring, often under acidic conditions to promote the requisite stereochemistry.^[39]^[40] Modern methods have expanded penam synthesis beyond classical routes, incorporating enzymatic and catalytic strategies for greater efficiency and diversity. Engineered δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetase, a non-ribosomal peptide synthetase, has been modified through site-directed mutagenesis to accept non-natural amino acid substrates, enabling the biosynthesis of novel ACV analogs that serve as precursors for penam derivatives via subsequent cyclization. Post-2000 advancements also include palladium-catalyzed cross-couplings, such as Sonogashira or Heck reactions on β-lactam halides, to functionalize the penam scaffold at the 2- or 3-position while preserving ring integrity.^[41]^[42] These approaches facilitate the synthesis of fluorinated or extended-side-chain analogs with improved pharmacological profiles. A primary challenge in penam synthesis remains maintaining the precise (2S,5R,6R) stereochemistry across the bicyclic system, as epimerization at C-5 or C-6 can occur during acidic or basic steps, often requiring chiral auxiliaries or asymmetric catalysis. Additionally, the inherent strain in the β-lactam ring predisposes it to nucleophilic opening by reagents or solvents, necessitating mild conditions and protective groups to achieve overall yields above 20% in total syntheses.^[43]^[44]

Chemical Properties

Stability and Reactivity

The β-lactam ring within the penam core exhibits significant ring strain, primarily due to bond angles of approximately 90° around the nitrogen and carbonyl carbon, which deviate markedly from the ideal tetrahedral angle of 109.5°.^[45] This strain enhances the electrophilicity of the carbonyl carbon, thereby increasing the susceptibility of penam to nucleophilic attack.^[46] The resulting heightened reactivity is a key chemical feature that distinguishes penam from larger lactams or linear amides, facilitating both its therapeutic action and potential for degradation.^[47] Hydrolysis of the penam β-lactam ring proceeds via nucleophilic attack at the carbonyl carbon by water (or other nucleophiles), leading to irreversible ring opening and formation of penicilloic acid as the primary degradation product.^[48] This process can be represented by the simplified equation:

\text{Penam} + \text{H}_2\text{O} \rightarrow \text{penicilloic acid (ring-opened carboxylic acid and amine)}

In the absence of enzymatic catalysis, the reaction is base-catalyzed under neutral or alkaline conditions but accelerates under acidic environments, where protonation of the carbonyl oxygen facilitates nucleophilic addition.^[49] While β-lactamases employ a similar mechanism enzymatically, the intrinsic chemical pathway underscores penam's vulnerability to aqueous environments.^[8] Stability of penam is notably pH-dependent, with pronounced acid lability at gastric pH levels (below 4), where the β-lactam ring undergoes rapid hydrolysis, limiting oral bioavailability of many derivatives.^[50] Conversely, stability improves in neutral to slightly basic media (pH 6–8), though prolonged exposure still leads to degradation. Substituents on the penam framework, particularly at the 6- and 7-positions, modulate this reactivity; electron-withdrawing side chains can sterically hinder nucleophilic access or electronically stabilize the carbonyl, enhancing resistance to hydrolysis in some analogs.^[46] Thermally, penam structures display conformational instability above 100°C, attributed to the puckered bicyclic geometry that promotes bond weakening and decarbonylation pathways upon heating.^[51] This leads to decomposition products such as penicillenic acid or further fragmented species, with vigorous gas evolution observed during melting or elevated-temperature processing.^[52] Such thermal sensitivity necessitates careful handling in pharmaceutical formulations to preserve integrity.

Spectroscopic Characteristics

Infrared (IR) spectroscopy reveals the β-lactam carbonyl stretch of penam at approximately 1780 cm⁻¹, notably higher than the 1660 cm⁻¹ typical for unstrained amides, owing to the ring strain that reduces resonance stabilization.^[6] This elevated frequency serves as a diagnostic feature for the strained four-membered ring in penam derivatives.^[53] Nuclear magnetic resonance (NMR) spectroscopy provides key insights into the penam core. In ¹H NMR, the methine protons at C-6 and C-7 exhibit chemical shifts in the range of 5.0-5.5 ppm, influenced by the deshielding effect of the adjacent β-lactam carbonyl and the fused ring system. For ¹³C NMR, the C-7 carbonyl carbon appears around 170 ppm, consistent with the partial amide character diminished by strain.^[54] X-ray crystallography elucidates the structural distortions in penam. The amide C-N bond measures 1.406 Å, elongated relative to standard amides (1.32 Å), while the C=O bond is shortened to 1.205 Å; these metrics reflect reduced π-overlap due to ring constraints.^[55] The bridgehead nitrogen displays pyramidal geometry, which limits planarity and contributes to reactivity.^[56] The thiazolidine ring adopts an envelope conformation, with cis-fusion between the β-lactam and five-membered rings.^[57] Mass spectrometry of penam cores or simple derivatives shows a molecular ion at m/z 129, corresponding to the protonated bicyclic fragment (C₅H₇NOS⁺); prominent fragments arise from β-lactam ring opening, such as loss of CO or cleavage at the amide bond, yielding ions indicative of the strained structure.^[58]

Biological Role

Role in Beta-Lactam Antibiotics

The penam nucleus serves as the defining core structure for penicillins, a major subclass of β-lactam antibiotics characterized by a β-lactam ring fused to a five-membered thiazolidine ring.^[59] Penicillins are naturally produced by certain filamentous fungi, primarily species of Penicillium such as P. chrysogenum and P. rubens, as well as Aspergillus nidulans.^[60] These compounds represent one of the earliest discovered classes of β-lactams, with penams forming the foundational scaffold that has enabled the development of a wide array of antimicrobial agents.^[59] In nature, penams function as secondary metabolites that provide ecological advantages to producing fungi, particularly in competitive microbial environments where they act as chemical defenses against bacterial competitors.^[32] For instance, Penicillium species secrete penicillins to inhibit the growth of surrounding bacteria, such as staphylococci, thereby securing resources like nutrients in soil or decaying organic matter.^[61] Several natural penicillins have been identified, with benzylpenicillin (penicillin G) and phenoxymethylpenicillin (penicillin V) serving as the primary examples; penicillin G is derived directly from Penicillium fermentation and exhibits activity against Gram-positive pathogens like Streptococcus species and Neisseria meningitidis, while penicillin V offers similar but orally bioavailable properties.^[59] Although dozens of natural penam variants have been isolated from fungal sources under varying conditions, semi-synthetic derivatives predominate in clinical applications due to enhanced stability and broader utility.^[62] Compared to other β-lactam cores, penams confer a relatively narrow antibacterial spectrum, primarily targeting Gram-positive bacteria and select Gram-negatives such as those causing syphilis or endocarditis, but with limited efficacy against Pseudomonas or anaerobes.^[59] In contrast, cephems (the core of cephalosporins) provide progressively broader coverage across generations, extending to respiratory and urinary tract pathogens, while carbapenems offer the widest spectrum, encompassing Gram-positive, Gram-negative, and anaerobic bacteria, including many resistant strains in nosocomial settings.^[59] This positions penams as foundational but specialized within the β-lactam family, with their natural occurrence underscoring the evolutionary innovation of fungal antibiotic production.^[32]

Mechanism of Action

Penam-based antibiotics, exemplified by penicillins, inhibit bacterial cell wall synthesis by mimicking the acyl-D-alanyl-D-alanine (D-Ala-D-Ala) terminus of the peptidoglycan precursor, which enables their recognition and binding by penicillin-binding proteins (PBPs).^[63] The bicyclic β-lactam ring system in the penam core provides the structural similarity to this substrate, facilitating initial reversible binding to the active site of PBPs, which are primarily transpeptidases responsible for cross-linking peptidoglycan strands during cell wall assembly.^[63]^[64] This binding triggers irreversible acylation, where the β-lactam carbonyl undergoes nucleophilic attack by the serine hydroxyl group in the PBP active site, leading to ring opening and formation of a stable acyl-enzyme complex.^[64]^[65] The reaction proceeds as follows:

\beta\text{-lactam ring} + \text{Ser-OH (PBP)} \rightarrow \text{penicilloyl-Ser (acyl-enzyme complex)}

This covalent modification inactivates the transpeptidase, preventing the cross-linking of peptidoglycan chains essential for cell wall integrity.^[64] The process unfolds in stages: initial non-covalent binding, followed by β-lactam ring opening via nucleophilic acylation by the PBP serine, stable adduct formation, and subsequent inhibition of peptidoglycan maturation, which weakens the cell wall and activates autolytic enzymes, ultimately causing bacterial lysis.^[65] Bacterial resistance to penam-based antibiotics arises through several mechanisms, including enzymatic hydrolysis by β-lactamases, which cleave the β-lactam ring to restore PBP functionality; active efflux pumps that reduce intracellular drug concentrations; and mutations in PBPs that decrease binding affinity.^[66]^[67] These antibiotics exhibit a spectrum primarily effective against Gram-positive bacteria, with variable efficacy against certain Gram-negative organisms due to barriers like the outer membrane that limit drug access to PBPs.^[68]^[66]

Derivatives and Applications

Penicillin Derivatives

Penicillin derivatives encompass a range of natural and semi-synthetic compounds built upon the penam core structure, primarily through variations in the side chain attached at the 6-amino position. These modifications enhance antibacterial spectrum, stability, and pharmacokinetic properties compared to the unmodified penam nucleus. Natural penicillins, produced directly via fungal fermentation, include Penicillin G, which features a benzyl side chain and was the first clinically used penicillin effective against gram-positive bacteria such as streptococci.^[69] Penicillin V, with a phenoxyacetyl side chain, offers improved oral bioavailability due to acid stability, making it suitable for outpatient treatment of similar infections.^[70] Penicillin F, characterized by a hexenoyl side chain, represents a minor natural variant with activity against select gram-positive organisms, though it is not widely used clinically.^[71] Semi-synthetic penicillins emerged by acylation of the 6-aminopenicillanic acid (6-APA) core with synthetic side chains, expanding therapeutic applications. Ampicillin, featuring an aminobenzyl side chain, was introduced in 1961 as the first broad-spectrum penicillin, extending efficacy to certain gram-negative bacteria like Escherichia coli while retaining gram-positive coverage.^[72] Amoxicillin, a hydroxylated variant of ampicillin, provides superior oral absorption and similar broad-spectrum activity, becoming a cornerstone for respiratory and urinary tract infections.^[72] Methicillin, with a 2,6-dimethoxyphenyl side chain, was developed in the late 1950s to combat staphylococcal penicillinases (beta-lactamases) and was effective against penicillin-resistant Staphylococcus aureus upon its 1960 introduction. However, methicillin-resistant S. aureus (MRSA) strains emerged shortly thereafter.^[72] Side-chain acylation at the 6-amino group of 6-APA allows tailored modifications to improve antibacterial spectrum, acid resistance, and beta-lactamase stability. For instance, amino-substituted chains in ampicillin and amoxicillin enhance gram-negative penetration, while bulky isoxazole rings in methicillin sterically hinder enzyme hydrolysis.^[73] These alterations address limitations of natural penicillins, such as narrow spectrum and poor oral uptake, without altering the core penam beta-lactam ring.^[74] Production of semi-synthetic penicillins typically involves fermentation of Penicillium chrysogenum to yield Penicillin G, followed by enzymatic hydrolysis using penicillin acylase to generate 6-APA, and subsequent chemical or enzymatic coupling of the desired side chain.^[75] This process, scaled industrially since the 1960s, enables efficient synthesis of variants like ampicillin from the common 6-APA intermediate.^[76] The 1960s marked a pivotal era for broad-spectrum penam derivatives, with the introduction of aminopenicillins like ampicillin and amoxicillin revolutionizing treatment of mixed infections and driving widespread antibiotic use.^[72] These developments, building on 6-APA isolation in 1957, shifted penicillin therapy from narrow gram-positive focus to versatile applications, though they also spurred resistance emergence.^[76]

Synthetic Analogs and Modifications

Synthetic analogs of the penam scaffold have been developed to expand the antimicrobial spectrum and overcome resistance mechanisms, primarily through structural modifications to the core bicyclic system. Penems represent a key class of unsaturated variants, featuring a double bond between C-2 and C-3 in the five-membered thiazolidine ring, which enhances their stability against certain β-lactamases and broadens activity against gram-negative bacteria compared to classical penams.^[46] These compounds are fully synthetic and include subclasses such as alkylpenems and arylpenems differentiated by substituents at the C-3 position. Carbapenem hybrids, such as imipenem, further modify the scaffold by replacing the sulfur atom with a carbon in the five-membered ring, resulting in potent activity against a wide range of pathogens, including anaerobes and Pseudomonas species, while maintaining the β-lactam core's mechanism of penicillin-binding protein inhibition.^[77] Imipenem's broader spectrum is attributed to its resistance to hydrolysis by many class A β-lactamases, though it remains susceptible to metallo-β-lactamases.^[78] A prominent modification involves oxidation of the sulfur atom to form 1,1-dioxo-penams, exemplified by sulbactam, which acts as a mechanism-based β-lactamase inhibitor rather than a direct antibiotic. Sulbactam, derived from penicillanic acid sulfone, irreversibly acylates the active site serine of class A β-lactamases, protecting partner β-lactams from degradation and restoring their efficacy against resistant strains.^[79] This sulfone group enhances the compound's stability and inhibitory potency, making it suitable for combination use.^[80] Post-2010 advances have focused on optimizing penem scaffolds for improved pharmacokinetics and activity against multidrug-resistant pathogens. Sulopenem, a thiopenem analog with a cyclic sulfoxide at C-3, was approved by the FDA in 2024 for uncomplicated urinary tract infections, marking the first oral penem available in the United States; its prodrug form, sulopenem etzadroxil, improves bioavailability over earlier penems like faropenem.^[81] Another development is the T405 series of penems, reported in 2020–2022, which demonstrate potent activity against nontuberculous mycobacteria such as Mycobacteroides abscessus, with derivatives like T422 and T428 showing enhanced efficacy through structural tweaks at the C-2 position.^[81] Fluorination of β-lactam cores has been explored to boost metabolic stability and resistance to enzymatic degradation; for instance, trifluoromethyl-substituted β-lactams exhibit improved antibacterial potency and reduced hydrolysis rates.^[82] Click chemistry has enabled the synthesis of targeted conjugates, such as boronic acid-based β-lactamase inhibitors identified via in situ azide-alkyne cycloadditions, enhancing specificity against resistant enzymes.^[83] As of 2025, ongoing research includes new penem derivatives with enhanced activity against resistant bacteria, as reviewed in recent studies.^[81] These analogs find applications in combination therapies to combat resistance, such as piperacillin-tazobactam, where the penam-derived piperacillin targets cell wall synthesis and tazobactam inhibits β-lactamases, providing broad coverage against gram-negative infections including those caused by extended-spectrum β-lactamase producers.^[84] Similarly, ampicillin-sulbactam pairs a classical penam with the 1,1-dioxo inhibitor for skin and respiratory infections.^[85] Imipenem is often combined with cilastatin to mitigate renal toxicity by inhibiting its metabolism.^[77] Despite these innovations, challenges persist in balancing enhanced activity with reduced toxicity and bioavailability. For example, carbapenems like imipenem can induce seizures and nephrotoxicity at high doses, necessitating careful dosing and combinations to minimize adverse effects while maintaining efficacy against resistant strains.^[77] Ongoing efforts aim to address efflux-mediated resistance and poor oral absorption in penem analogs through further scaffold hybridization.^[81]

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This year (2007) marks the 50th anniversary of [corrected] discovery of 6-aminopenicillanic acid (6-APA), the precursor of all semi-synthetic penicillins.Missing: isolation 1945
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The 50th anniversary of the discovery of 6-aminopenicillanic acid (6 ...
Aug 5, 2025 · This year (2007) marks the 50th anniversary of the discovery of 6-aminopenicillanic acid (6-APA), the precursor of all semi-synthetic penicillins and ...
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[PDF] The chemical structure of the penicillins - Nobel Prize
Thus, the structure of the thiocyanic degradation product of penicillin is really against structure (X), though an intramolecular re-. Page 32. CHEMICAL ...Missing: 1940s | Show results with:1940s
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The chemical structure of penicillin was determined between 1942 and 1945 under conditions of secrecy established by the U.S. and U.K. governments.
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[PDF] Dorothy Crowfoot Hodgkin - Nobel Lecture
Here it is easy to see the chemical structure, a four-membered ring attached to a six-membered sulphur-containing ring-derived most prob- ably like the ...
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Molecular Control of Expression of Penicillin Biosynthesis Genes in ...
Penicillin biosynthesis in P. · High glucose concentrations prevent formation of ACV (the first intermediate of the pathway) and depress isopenicillin N synthase ...
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The cluster of penicillin biosynthetic genes. Identification and ...
Penicillium chrysogenum DNA fragments cloned in EMBL3 or cosmid vectors from the upstream region of the pcbC-penDE cluster carry a gene (pcbAB) that ...
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The cluster of penicillin biosynthetic genes. Identification ... - PubMed
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Isopenicillin N synthetase of Penicillium chrysogenum, an enzyme ...
Isopenicillin N Synthetase of Penicillium chrysogenum,an Enzyme. That Converts 8-(L-oL-Aminoadipyl)-L-Cysteinyl-D-Valine to. Isopenicillin N.
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Penicillin production in industrial strain Penicillium chrysogenum ...
Feb 16, 2017 · The three penicillin biosynthesis genes pcbAB, pcbC, and penDE are clustered in a single 18-kb region in wild-type strains of the filamentous ...
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Penicillin Biosynthesis - News-Medical.net
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Increased Penicillin Production in Penicillium chrysogenum ...
Sep 7, 2012 · The penicillin biosynthesis cluster in P. chrysogenum consists of three genes, pcbAB, pcbC, and penDE (Fig. 1A), encoding the enzymes that ...
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Transcription of the pcbAB, pcbC and penDE genes of Penicillium ...
Summary: Glucose repressed transcription of the penicillin biosynthesis genes pcbAB, pcbC and penDE when added at inoculation time to cultures of Penicillium ...
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Comparative genomics of Alexander Fleming's original Penicillium ...
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Molecular biology of penicillin and cephalosporin biosynthesis
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All semi-synthetic penicillins manufactured starting from 6-aminopenicillanic acid (6-APA) are produced through a fermentation process.
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Use of enzymes in the production of semi-synthetic penicillins and ...
In fact, the hydrolysis of penicillin G to produce 6-APA catalyzed by penicillin G acylase is one of the most successful historical examples of the enzymatic ...
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Preparation of β-lactams by [2+2] cycloaddition of ketenes and imines
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The Chemical Relationship Among Beta-Lactam Antibiotics and ...
Penams are a class of β-lactams possessing an azabicyclo[3.2.0]heptane ring system, a carboxylic acid moiety at C3, and a sulfur atom at position one (Figure 1) ...Missing: stereochemistry | Show results with:stereochemistry<|control11|><|separator|>
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The Chemical Relationship Among Beta-Lactam Antibiotics and ...
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[PDF] Improvements of Amoxicillin Stability in Acidic Environment
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A Thermal Decarbonylation of Penam β-Lactams - PMC
When 7 was heated on a hot stage (mp 95-98 °C), vigorous bubbling was observed above 100 °C and had finished when the sample had reached 155 °C ...
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(PDF) Solid state thermal and spectroscopic studies on the antibiotic ...
Aug 10, 2025 · At temperatures higher than 205 °C the thermal decomposition took place with elimination of methylamine in agreement with the first mass ...
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Linear and Non-Linear Middle Infrared Spectra of Penicillin G in the ...
Jan 8, 2021 · The band at 1767 cm − 1 is associated with the stretching of the CO bond of the β − l a c t a m , while that at 1674 cm − 1 to Amide I mode and ...
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13 C NMR characteristics of penicillins. 13 C NMR Chemical Shifts of...
The carbonyl group in the ring, due to the non-planarity, is not conjugated resulting in strong electron deficiency at the carbonyl carbon (Szabó et al., 2015) ...Missing: 1H H-
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Attaching Metal-Containing Moieties to β-Lactam Antibiotics
Jun 26, 2024 · This is the first structural report of a penicillin-transition-metal complex having the bicyclic system of these antibiotics intact.
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Stereospecific synthesis, 1H and 13C NMR spectroscopy, and X-ray ...
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Chemical-ionization mass spectrometry of beta-lactam antibiotics
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... commercial production. The significance of the identification of 6-APA is that numerous semisynthetic penicillins can be prepared by adding acyl side-chain ...
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The penicillin-binding proteins: structure and role in peptidoglycan ...
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Direct enzymatic synthesis of natural penicillins using phenylacetyl ...
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New Semisynthetic Penicillins Obtained by Coupling of the 6 ... - NIH
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6-APA can be considered the most important intermediate in the synthesis of both semisynthetic and naturally occurring penicillins.
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Process Development for 6-Aminopenicillanic Acid Production ...
Nov 6, 2020 · Immobilized penicillin acylase is a biocatalyst suitable for the kinetically controlled industrial synthesis of semi-synthetic antibiotics in aq ...
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The 50th anniversary of the discovery of 6-aminopenicillanic acid (6 ...
Production of 6-APA by enzymatic hydrolysis of penicillin. Notwithstanding the inability to confirm the work of Sakaguchi and Murao, a programme of work was ...
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The discovery of penicillin in 1929 [1] is rightly recognized as a milestone in the history of medicine, and its introduction to the clinic in the 1940s ...
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HU205611B - Google Patents
Sulbactam (penicillanic acid 1,1-dioxide) or pharmaceutically acceptable salts thereof are well known in the art as β-lactamase inhibiting antibiotics.
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Recent Developments in Penem Antibiotics: Structural and ...
May 11, 2025 · Similar to β-lactam antibiotics, penems inhibit PBPs, which results in bactericidal action [1]. Penems can be obtained by total synthesis or ...
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Synthesis and Antibacterial Activity of Novel Phosphonated CF3-β ...
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Discovery of Boronic Acids-Based β-Lactamase Inhibitors Through ...
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Resistance to β-lactams is primarily because of bacterially produced β-lactamase enzymes that hydrolyze the β-lactam ring, thereby inactivating the drug.
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Sulbactam-containing beta-lactamase inhibitor combinations
Sulbactam irreversibly inhibits the hydrolytic activity of beta-lactamases. This compound is commercially available in combination with either ampicillin or ...